Objective:To investigate the role and underlying mechanism of protein arginine methyltransferase 1 (PRMT1) and its inhibitor in alkali burn-induced corneal neovascularization (CNV).Methods:Seventy-two SPF-grade C57BL/6 mice were randomly divided into a normal group and 1 day post-modeling, 4 days post-modeling, and 7 days post-modeling groups to establish an alkali burn-induced CNV model and determine the optimal time point for analysis.Another 90 mice were randomly assigned to five groups: alkali burn group, dimethyl sulfoxide (DMSO) group, PRMT1 inhibitor group, fibroblast growth factor 2 (FGF2) inhibitor group, and PRMT1 inhibitor combined with FGF2 group to evaluate the role of PRMT1 in CNV.Human umbilical vein endothelial cells (HUVECs) and murine macrophage-like RAW264.7 cells were used to establish a hypoxia/reoxygenation (H/R)-induced in vitro model to mimic the ischemic microenvironment.Cells were assigned to the following groups: control group, H/R group, H/R+ DMSO group, H/R+ si-NC group, H/R+ si-PRMT1 group, H/R+ si-FGF2 group, H/R+ PRMT1 inhibitor group, and H/R+ PRMT1 inhibitor+ FGF2 group.Corneal opacity and CNV areas were assessed by slit-lamp microscopy.Corneal structural changes and inflammatory cell count were determined by hematoxylin and eosin staining.PRMT1-positive cell count was determined by immunohistochemistry and the expression of PRMT1, CD31, vascular endothelial growth factor (VEGF), F4/80, CD206, and inducible nitric oxide synthase (iNOS) was assessed by immunofluorescence staining.The expression levels of macrophage markers, including F4/80, iNOS, CD206, interleukin-10 (IL-10), and arginase-1 (Arg-1), were quantified by real-time quantitative PCR and Western blot.Cell proliferation, migration, and angiogenic capacity were evaluated by functional assays including the CCK-8 assay, wound healing assay, Transwell migration assay, and tube formation assay.The research process followed the relevant regulations of the Visual and Ophthalmology Association, and the research plan was approved by the Laboratory Animal Committee of Wuhan University (No.20220504A). Results:Compared with the normal group, the 7 days post-modeling group showed significantly increased corneal opacity scores and CNV area, upregulated VEGF expression, and increased inflammatory cells (all P<0.05).The number of PRMT1-positive cells in the alkali burn group was (39.67±3.51) cells/visual field, which was significantly higher than (3.33±0.58) cells/visual field in the normal group ( t=17.68, P<0.01).Both mRNA and protein expression levels of PRMT1 and FGF2 were significantly elevated in the alkali burn group compared with the normal group (all P<0.01).Compared with the alkali burn group, the PRMT1 inhibitor group showed reduced corneal opacity scores, decreased CNV area, fewer inflammatory cells, and lower expression levels of PRMT1, FGF2, VEGF, Arg-1, IL-10 proteins, as well as CD206 mRNA (all P<0.05).Cell viability, migration distance, migration number, and tubes formed were significantly increased in the H/R group compared with the control group, significantly reduced in the H/R+ si-PRMT1 and H/R+ PRMT1 inhibitor groups compared with the H/R group and significantly increased in H/R+ PRMT1 inhibitor+ FGF2 group than in H/R+ PRMT1 inhibitor group (all P<0.05).Compared with the H/R group, the H/R+ PRMT1 inhibitor group exhibited reduced expression of FGF2, VEGFA, p-PI3K, and p-Akt, while those were upregulated in the H/R+ PRMT1 inhibitor+ FGF2 group compared with the H/R+ PRMT1 inhibitor group (all P<0.05).The proportions of CD206-positive cells in the H/R, H/R+ DMSO, H/R+ PRMT1 inhibitor, and H/R+ PRMT1 inhibitor+ FGF2 groups were all significantly higher than those in the control group, and significantly higher in the H/R, H/R+ DMSO, and H/R+ PRMT1 inhibitor+ FGF2 groups compared with the H/R+ PRMT1 inhibitor group (all P<0.05).Compared with the alkali burn group, the FGF2 inhibitor group, PRMT1 inhibitor group, and PRMT1 inhibitor+ FGF2 group all showed reduced corneal opacity scores, CNV area, and decreased number of VEGFA-, CD206-, and F4/80-positive cells, with the above indicators being lower in the PRMT1 inhibitor group compared with the FGF2 inhibitor and PRMT1 inhibitor+ FGF2 groups and higher in PRMT1 inhibitor+ FGF2 group than in the FGF2 inhibitor group (all P<0.05).Compared with the alkali burn group, the PRMT1 inhibitor group had decreased protein expression levels of FGF2, p-PI3K, p-Akt, CD31, VEGFA and Arg-1, with higher protein expression levels in the PRMT1 inhibitor+ FGF2 group than in the PRMT1 inhibitor group (all P<0.05). Conclusions:PRMT1 may regulate macrophage activation and anti-inflammatory polarization via the FGF2/PI3K/Akt signaling pathway, thereby promoting the occurrence and development of CNV.Targeted inhibition of PRMT1 may serve as an effective therapeutic strategy for CNV.

Objective:To investigate the role and underlying mechanism of protein arginine methyltransferase 1 (PRMT1) and its inhibitor in alkali burn-induced corneal neovascularization (CNV).Methods:Seventy-two SPF-grade C57BL/6 mice were randomly divided into a normal group and 1 day post-modeling, 4 days post-modeling, and 7 days post-modeling groups to establish an alkali burn-induced CNV model and determine the optimal time point for analysis.Another 90 mice were randomly assigned to five groups: alkali burn group, dimethyl sulfoxide (DMSO) group, PRMT1 inhibitor group, fibroblast growth factor 2 (FGF2) inhibitor group, and PRMT1 inhibitor combined with FGF2 group to evaluate the role of PRMT1 in CNV.Human umbilical vein endothelial cells (HUVECs) and murine macrophage-like RAW264.7 cells were used to establish a hypoxia/reoxygenation (H/R)-induced in vitro model to mimic the ischemic microenvironment.Cells were assigned to the following groups: control group, H/R group, H/R+ DMSO group, H/R+ si-NC group, H/R+ si-PRMT1 group, H/R+ si-FGF2 group, H/R+ PRMT1 inhibitor group, and H/R+ PRMT1 inhibitor+ FGF2 group.Corneal opacity and CNV areas were assessed by slit-lamp microscopy.Corneal structural changes and inflammatory cell count were determined by hematoxylin and eosin staining.PRMT1-positive cell count was determined by immunohistochemistry and the expression of PRMT1, CD31, vascular endothelial growth factor (VEGF), F4/80, CD206, and inducible nitric oxide synthase (iNOS) was assessed by immunofluorescence staining.The expression levels of macrophage markers, including F4/80, iNOS, CD206, interleukin-10 (IL-10), and arginase-1 (Arg-1), were quantified by real-time quantitative PCR and Western blot.Cell proliferation, migration, and angiogenic capacity were evaluated by functional assays including the CCK-8 assay, wound healing assay, Transwell migration assay, and tube formation assay.The research process followed the relevant regulations of the Visual and Ophthalmology Association, and the research plan was approved by the Laboratory Animal Committee of Wuhan University (No.20220504A). Results:Compared with the normal group, the 7 days post-modeling group showed significantly increased corneal opacity scores and CNV area, upregulated VEGF expression, and increased inflammatory cells (all P<0.05).The number of PRMT1-positive cells in the alkali burn group was (39.67±3.51) cells/visual field, which was significantly higher than (3.33±0.58) cells/visual field in the normal group ( t=17.68, P<0.01).Both mRNA and protein expression levels of PRMT1 and FGF2 were significantly elevated in the alkali burn group compared with the normal group (all P<0.01).Compared with the alkali burn group, the PRMT1 inhibitor group showed reduced corneal opacity scores, decreased CNV area, fewer inflammatory cells, and lower expression levels of PRMT1, FGF2, VEGF, Arg-1, IL-10 proteins, as well as CD206 mRNA (all P<0.05).Cell viability, migration distance, migration number, and tubes formed were significantly increased in the H/R group compared with the control group, significantly reduced in the H/R+ si-PRMT1 and H/R+ PRMT1 inhibitor groups compared with the H/R group and significantly increased in H/R+ PRMT1 inhibitor+ FGF2 group than in H/R+ PRMT1 inhibitor group (all P<0.05).Compared with the H/R group, the H/R+ PRMT1 inhibitor group exhibited reduced expression of FGF2, VEGFA, p-PI3K, and p-Akt, while those were upregulated in the H/R+ PRMT1 inhibitor+ FGF2 group compared with the H/R+ PRMT1 inhibitor group (all P<0.05).The proportions of CD206-positive cells in the H/R, H/R+ DMSO, H/R+ PRMT1 inhibitor, and H/R+ PRMT1 inhibitor+ FGF2 groups were all significantly higher than those in the control group, and significantly higher in the H/R, H/R+ DMSO, and H/R+ PRMT1 inhibitor+ FGF2 groups compared with the H/R+ PRMT1 inhibitor group (all P<0.05).Compared with the alkali burn group, the FGF2 inhibitor group, PRMT1 inhibitor group, and PRMT1 inhibitor+ FGF2 group all showed reduced corneal opacity scores, CNV area, and decreased number of VEGFA-, CD206-, and F4/80-positive cells, with the above indicators being lower in the PRMT1 inhibitor group compared with the FGF2 inhibitor and PRMT1 inhibitor+ FGF2 groups and higher in PRMT1 inhibitor+ FGF2 group than in the FGF2 inhibitor group (all P<0.05).Compared with the alkali burn group, the PRMT1 inhibitor group had decreased protein expression levels of FGF2, p-PI3K, p-Akt, CD31, VEGFA and Arg-1, with higher protein expression levels in the PRMT1 inhibitor+ FGF2 group than in the PRMT1 inhibitor group (all P<0.05). Conclusions:PRMT1 may regulate macrophage activation and anti-inflammatory polarization via the FGF2/PI3K/Akt signaling pathway, thereby promoting the occurrence and development of CNV.Targeted inhibition of PRMT1 may serve as an effective therapeutic strategy for CNV.

Purpose/Significance To construct a specialized database for inflammatory demyelinating disease of the central nervous system(CNS),so as to contribute to clinical research and improve the diagnostic and treatment capabilities of primary healthcare institu-tions.Method/Process Using the internet to collect medical data,after processing and analysis,the CNS inflammatory demyelinating disease database is constructed.Using statistical analysis,natural language processing(NLP),artificial intelligence(AI)image recog-nition and data visualization and other technologies,the database information is integrated and analyzed.Result/Conclusion A standard-ized big database for CNS inflammatory demyelinating diseases is constructed,which enables visualization of clinical research data,pro-vides patient education and specialist training,and facilitates multi-center teleconsultations.The establishment of a specialized database for the CNS inflammatory demyelinating disease can promote the transformation of medical research achievements,provide references for future real-world clinical research,optimize the process of diagnosis and treatment,and improve the clinical capability of primary healthcare institutions.

Objective: To evaluate the role of hippocampal mast cells in the early postoperative cognitive impairment in rats. Methods: Eighty male Sprague-Dawley rats, aged 6-8 weeks, weighing 200-250 g, were divided into 4 groups (n=20 each) using a random number table method: normal saline control group (group C), cromolyn injected into lateral cerebral ventricle group (group L), operation group (group O), and cromolyn injected into lateral cerebral ventricle plus operation group (group LO). Cromolyn 2 μl (100 μg/ul) was intracerebroventricularly injected in L and LO groups.The equal volume of normal saline was given instead in C and O groups.Open reduction and internal fixation was performed after tibial fracture was induced 30 min later in O and LO groups.Contextual fear conditioning and Y-maze tests were performed at 1 and 3 days after operation, and the freezing time and the number of learning trails were recorded.The animals were then sacrificed, and hippocampal tissues were obtained for determination of activated mast cell count (by toluidine blue staining) and expression of tumor necrosis factor-alpha (TNF-α) and interleukin-1beta (IL-1β) mRNA (by polymerase chain reaction). Results: Compared with group C, the number of learning trails was significantly increased, and the freezing time was shortened, the number of activated mast cells in hippocampi was increased, and the expression of IL-1β mRNA was up-regulated at 1 and 3 days after operation, and the expression of TNF-α mRNA was up-regulated at 1 day after operation in group O(P<0.05), and no significant change was found in the parameters mentioned above in group L (P>0.05). Compared with group O, the number of learning trails was significantly decreased, and the freezing time was prolonged, the number of activated mast cells in hippocampi was decreased, and the expression of IL-1β mRNA in hippocampi was down-regulated at 1 and 3 days after operation, and the expression of TNF-α mRNA in hippocampi was down-regulated at 1 day after operation in group LO (P<0.05). Conclusion Activation of hippocampal mast cells can induce central inflammatory responses and is involved in the mechanism of early postoperative cognitive impairment in rats.

To explore the effect of propofol on human cardiac AC16 cells under CoCl2-induced hypoxic injury and the possible mechanisms.
 Methods: Human AC16 cardiomyocytes were treated with cobalt chloride (CoCl2) to mimic hypoxic condition in cultured cardiomyocytes. The AC16 cells were divided into 3 groups: a control group, a CoCl2 hypoxia group (CoCl2 group), and a propofol+CoCl2 group (propofol+ CoCl2 group). The cell viability was assessed by cell counting kit-8 (CCK-8). Cell apoptosis ratio (AR) and the mitochondrial membrane potential (Δψm) were detected by flow cytometry. The reactive oxygen species (ROS) production in AC16 cells were determined with the ROS-sensitive fluorescent probe. Meanwhile, total intracellular levels of malondialdehyde (MDA) and superoxide dismutase (SOD) in AC16 cells were detected with commercially available kits. Western blot was used to evaluate the activation of c-Jun N-terminal kinase (JNK) and p38 signaling pathways.
 Results: 1) Compared with the control group, AC16 cell viability was decreased significantly in the CoCl2 group following the treatment with 500 μmol/L CoCl2 (P<0.01); 2) Compared with the control group, AR value in AC16 cells was increased significantly in the CoCl2 group, while Δψm was decreased significantly (all P<0.01). Compared with the CoCl2 group, AR value in AC16 cells was decreased significantly in the propofol+CoCl2 group, while Δψm was increased significantly (both P<0.05); 3) Compared with the control group, the levels of ROS and MDA were increased significantly, and the level of SOD was significantly decreased in the CoCl2 group (all P<0.01). Compared with the CoCl2 group, the ROS and MDA levels in the propofol+CoCl2 group were increased significantly and the SOD levels were decreased significantly (all P<0.05); 4) Compared with the control group, the phosphorylation levels of JNK and p38 were increased significantly (both P<0.05) in the CoCl2 group. Compared with the CoCl2 group, the phosphorylation levels of JNK and p38 were decreased significantly in the propofol+CoCl2 group (both P<0.05).
 Conclusion: The pretreatment with propofol may protect human cardiac AC16 cells from the chemical hypoxia-induced injury through regulation of JNK and p38 signaling pathways.
Apoptosis ; Cell Hypoxia ; Cell Line ; Cell Survival ; Cobalt ; pharmacology ; Humans ; Hypoxia ; JNK Mitogen-Activated Protein Kinases ; Propofol ; Reactive Oxygen Species

Objective To investigate different priming dose of cisatracurium effect on the onset time.Methods In the First Affiliated Hospital of Nanjing Medical University from November 2014 to April 2015,eighty adult patients (male 41 cases,female 39 cases age from 18 to 60.) scheduled for selective surgery,were randomly divided into four groups,20 in each.Control group (group C) priming with 3 ml saline,group C1 priming with cisatracurium 15 μg/kg,group C2 priming with cisatra curium 30 μg/kg and group C3 priming with cisatracurium 50 μg/kg,1 minutes after the priming injection,each group respectively received the left over intubation dose of cisatracurium 0.15,0.135,0.12,0.10 mg/kg,followed by anesthesia induction of midazolam 0.05 mg/kg,fentanyl 5.0 μg/kg,etomidate 0.3 mg/kg.Neuromuscular block was monitored using train of four stimulation mode.The time when T4/T1=0 after the left over intubation dose of cisatracurium injection and adverse reaction were recorded.Results The onset time in group C3 (114.2±14.1) s was significantly less than that in group C2 (136.3±428.1) s,group C1 (164.6±26.9) s and group C (165.9±10.8) s (P＜0.01).No adverse reaction of dyspnea,urticaria,arrhythmia occurred after priming injection of cisatracurium in all the four groups.Conclusion Priming dose of 50 μg/mg cisatracurium can significantly shorten the onset time compared to the priming dose of 15 μg/mg and 30 μg/mg.

The present study was to examine the effect of stellate ganglion block (SGB) on bilateral regional cerebral oxygen saturation (rSO2) and postoperative cognitive function. Eighty patients undergoing selective coronary artery bypass graft with cardiopulmonary bypass (CPB) were randomly and equally divided into two groups. The patients in group S were given right SGB with ropivacaine, while the patients in group C were injected with normal saline. We compared the bilateral rSO2 after SGB. Minimum Mental State Examination (MMSE), Visual Verbal Learning Test (VVLT), and Digital Span Test (DST) were applied to observe the effect on cognitive function. We found that the incidence of postoperative cognitive dysfunction (POCD) 7 days after surgery in group S was lower than that in group C. The level of blocked side rSO₂ of S group were significantly higher before CPB time of rewarming than that before SGB (P < 0.05), much higher than corresponding non-blocked side rSO₂ before CPB (P < 0.05), and much higher than rSO₂ level in group C before CPB and after CPB (P < 0.05). The non-blocked side rSO₂ in group S before anesthesia were much lower than basic levels and those in group C (P < 0.05). It could be concluded from the above results that there was significant increase in the blocked-side rSO₂ compared to the non-blocked side and there was significant decrease in the incidence of POCD compared to the control group after SGB.
Autonomic Nerve Block ; adverse effects ; Cardiopulmonary Bypass ; adverse effects ; Cerebrum ; physiology ; Cognition ; Cognition Disorders ; Coronary Artery Bypass ; adverse effects ; Humans ; Incidence ; Oxygen ; physiology ; Oxygen Consumption ; Postoperative Complications ; Stellate Ganglion

Objective To observe the effect of creatine phosphate sodium on BIS and recovery quality during general anesthesia emergence period in elderly patients.Methods Sixty ASA Ⅰ or Ⅱpatients,31 males,29 females,aged 65-80 yr undergoing transabdominal cholecystectomy by general anesthesia were randomly allocated into two groups(n=30 each):group creatine phosphate (P)and group control(C)according to random numbers generated by computer.Patients were intravenously infused with 1.0 g creatine phosphate sodium melted in 100 ml normal saline or only 100 ml normal saline in group P or C respectively in thirty minutes at the same time of surgical incision.The heart rate(HR)and bispectral index(BIS)were recorded before anesthesia induction (T0 ),during sputum aspiration (T0 ),during extubation (T2 )and 1(T3 ),5(T4 ),10(T5 ),15 minutes (T6 )after extubation. The dosage of propofol,remifentanil and cisatracurium,anesthesia duration,operation time,awake time,extubation time,recovery time of consciousness and Steward recovery scores on T3-T6 were also recorded,and the occurrence of tachycardia during the operation was observed at the same time. Results Compared with T0 ,the BIS value were lower significantly and HR were significantly in-creased on T1-T4 in the two groups(P <0.05).Compared with group C,the BIS value were signifi-cantly higher in group P on T1-T4 (P <0.05).Compared with group C,awake time,extubation time, recovery time of consciousness significantly shortened in group P (P <0.05).There were six cases of tachycardia occurring in group C which were significantly higher than two cases in group P (P <0.05).Steward recovery scores on T3 and T4 were also higher in group P (P <0.05).Conclusion Not only can 1.0 g creatine phosphate sodium administered during transabdominal cholecystectomy im-prove the BIS value of general anesthesia recovery period and recovery quality,but also effectively re-duce the incidence rate of tachycardia during operation in elderly patients.

Objective To investigate the effect of IL-1 7A on lipopolysaccharide (LPS)-induced neuroinflammation and fear conditioning test.Methods Among 70 male SD rats aged 18 months, firstly,thirty rats were randomly divided into five groups:control group(group A),LPS 6 h group (group B),12 h group(group C),24 h group(group D),48 h group(group E).Group A were injected intraperitoneally with normal saline and groups B,C,D and E were injected with LPS 500 μg/kg.Ani-mals of groups A,B,C,D and E were killed respectively after LPS injection and their hippoeampus tis-sue was detected for the concentration of IL-1 7A.Secondly,forty rats were randomly divided into 4 groups:control group(group O),IL-1 7A antibody group(group P),LPS group (group Q),IL-1 7A antibody+LPS group(group R).Group P and group R were injected intracerebroventricularly with IL-1 7A antibodies 3 μl (200 μg/μl),groups O and Q were injected equal volume of normal saline.30 min later,groups Q and R were injected intraperitoneally with LPS 500 μg/kg,groups O and P were in-jected equal volume of normal saline.24 h later,contextual fear conditioning test was performed. Then,all animals were killed and their hippocampus tissue would be detected for the concentration of TNF-αand IL-6,as well as the expression of Iba1-positive cells.Results The concentration of IL-1 7A of groups B,C,D and E increased significantly compared with group A (P <0.01 ),there was no difference between groups E and A.The freezing time of groups Q and R was significantly shortened than that in group O(P <0.01 or P <0.05 ),the freezing time of group R was significantly longer than that in group Q(P <0.01).The concentration of TNF-αand IL-6 of groups Q and R was obvi-ously higher than group O(P <0.01 ),the concentration of TNF-α and IL-6 of group R lower than group Q(P <0.01).The expression of Iba1-positive cells in hippocampal area CA1 of groups Q and R was obviously increased compared with group O(P <0.01).Compared with group Q,the expression of Iba1-positive cells in hippocampal area CA1 of group R were obviously decreased (P < 0.05 ). Conclusion IL-1 7A is implicated in the early stage of LPS-induced neuroinflammation and the chan-ging of freezing time in contextual fear conditioning in aged rats.

Objective To improve the understanding of cutaneous intravascular natural killer/T-cell lymphoma (CIVNKTC). Methods Clinical data on five cases of CIVNKTC were collected. The histopathological feature, treatment and prognosis of CIVNKTC were retrospectively analyzed and discussed. Results Of the 5 patients, 1 was male and 4 were female. The age of onset ranged from 38 to 83 years (average, 56.2 years). All the patients presented with multiple plaques and nodules as the primary symptoms. Histopathological examination revealed vasodilatation in the dermis and subcutaneous tissue, as well as atypical lymphoid cells with large hyperchromatic nuclei containing 1-2 small nucleoli in dilated veins. Immunohistochemical studies of tumor cells showed positive staining for CD3ε, cytotoxic proteins (including T cell-restricted intracellular antigen-1, granzyme B and perforin)and Epstein-Barr virus(EBV)-encoded microRNA, but negative staining for cytokeratin, CD20, CD79a, CD4 and CD8. Furthermore, the tumor cells stained positive for CD56 in two patients. Among the 5 patients, only 2 received chemotherapy and the remaining received no treatment. During a 24-month follow-up, 4 patients died, and only 1 survived with the tumor. Conclusion CIVNKTC is a rare extranodal Hodgkin′s lymphoma with distinct histologic manifestations and immunophenotypes, rapid and aggressive clinical course, and poor prognosis.