Objective:To evaluate the role of ten-eleven translocation methylcytosine dioxygenase 3 (TET3) in trigeminal ganglion in maxillofacial inflammatory pain in mice.Methods:Forty SPF healthy male C57BL/6J mice, aged 8-10 weeks, weighing 19-23 g, were divided into 5 groups ( n=8 each) using a random number table method: control group (group C), inflammatory pain group (group IP), control+ TET3-siRNA group (group C+ siTET3), inflammatory pain+ TET3-siRNA group (group IP+ siTET3) and inflammatory pain+ negative control Scrambled-siRNA group (group IP+ siNC). Normal saline or complete Freund′s adjuvant (CFA) 10 μl was injected into the temporomandibular joint of mice, respectively, and the mechanical paw withdrawal threshold (MWT) was measured at 1, 4, 8 and 12 days after injection (T 1-4). Before injection of normal saline or CFA, 0.75 μl siTET3 or siNC was injected into the trigeminal ganglion and the animals were then sacrificed and trigeminal ganglion was removed at T 2 for determination of the expression of TET3 by Western blot in C+ siTET3, IP+ siTET3 and IP+ siNC groups. Results:Compared with group C, MWT was significantly decreased at T 1-3 , the expression of TET3 in trigeminal ganglion was up-regulate in group IP ( P<0.05 or 0.01). Compared with IP and IP+ siNC groups, MWT was significantly increased at T 2, 3, and the expression of TET3 in trigeminal ganglion was down-regulate in group IP+ siTET3 ( P<0.05 or 0.01). Conclusion:TET3 in trigeminal ganglion is involved in the development of maxillofacial inflammatory pain in mice.

Objective:To explore the correlation between the expression of signaling lymphocyte activation molecule family 6 (SLAMF6) on peripheral blood CD8 +T cells and perforin and granzyme B and the clinical significance in patients with newly diagnosed severe aplastic anemia(SAA). Methods:The indicators of blood routine and bone marrow and peripheral blood samples of 32 newly diagnosed SAA patients admitted to Henan Provincial People′s Hospital from January 2016 to June 2019 were collected for retrospective analysis. Flow cytometry was used to detect the expression of SLAMF6, perforin and granzyme B on samples CD8 +T cell before therapy and 6 months after therapy (11 cases received transplantation, 21 cases received immunosuppressive therapy [IST]). Spearman correlation analysis was performed to determine the association between clinical indicators and laboratory test results. The expression of SLAMF6, perforin and granzyme B was also detected in 10 healthy people (normal group) and 13 myelodysplastic syndromes/paroxysmal nocturnal hemoglobinuria (MDS/PNH) patients (MDS/PNH group). Results:(1) At diagnosis: the expression of SLAMF6 was significantly lower in the SAA group than that in the normal group and the MDS/PNH group ([56.40±6.37]% vs [84.34±5.81]% and [82.24±4.98]% (both P<0.001]). The expression of perforin was significantly higher in the SAA group (32.73±8.46) than that in the normal control group (23.75%±5.10%), and the MDS/PNH group (26.12%±5.53%) (both P<0.05). The expression of granzyme B was also significantly higher in the SAA group (36.23%±7.94%) than that in the normal control group (21.67%±5.05%) and the MDS/PNH group (21.79%±5.10%) (both P<0.001). The expression of SLAMF6 was positively correlated with the hemoglobin ( r=0.804), and reticulocyte absolute values ( r=0.656) in peripheral blood, percentage of granulocytes ( r=0.643) and erythrocytes ( r=0.622) in bone marrow of SAA patients (all P<0.05). Expression of SLAMF6 was negatively correlated with perforin ( r=-0.792) and granzyme B ( r=-0.908) on CD8 +T cells in patients with SAA (both P<0.001). (2) After treatment: the expression of SLAMF6 in peripheral blood CD8 +T cells of 30 surviving patients was higher than pre-treatment ([79.19±12.69]% vs [56.40±6.37]%, P<0.001). The expressions of perforin and granzyme B were lower than pre-treatment level (both P<0.05). The expression of SLAMF6 on CD8 +T cells in 11 transplanted patients was higher than before transplantation ([86.54±3.75]% vs [56.40±7.35]%, P<0.001). The expressions of perforin and granzyme B were lower than before transplantation (both P<0.05). The expression of SLAMF6 on CD8 +T cells in 12 IST-respond patients was higher than that before treatment, while the perforin and granzyme B levels were lower than pre-treatment (all P<0.05). The post-treatment expressions of SLAMF6, perforin and granzyme B were similar as before treatment levels in 7 IST-unrespond patients (all P>0.05). Conclusion:SLAMF6 is significantly down-regulated on CD8 +T cells in newly diagnosed SAA, negatively correlated with the effective factors of CD8 +T cells, which might participate in the immune regulatory of CD8 +T cells as a negative regulatory factor in patients with SAA. The SLAMF6 is significantly up-regulated after hematopoietic recovery, while there is no significant change in treatment-unrespond patients, which could thus serve as an useful diagnostic and therapeutic index of patients with SAA.

Objective:To investigate the neuroprotective effect of long-term prophylactic use of buphthalein on mice with permanent distal middle cerebral artery occlusion and its relationship with the nuclear factor erysid 2 related factor 2 (Nrf2) pathway.Methods:Nrf2 + /+ wild-type and Nrf2 -/- knockout mice were randomly divided into control group (equal volume vegetable oil), low-dose butylphthalide group (20 mg/kg) and high-dose butylphthalide group (60 mg/kg), with 6 mice in each group. The drug was administered once a day by gavage for 1 month, and then a permanent middle cerebral artery occlusion model was induced by electrocoagulation. After the model was made, the drug was continued and the mice were sacrificed on the 10 th day. The modified Longa grading scale and the rotating rod test were used to evaluate neurological deficits on the 3 rd and 10 th day after the model was made. After the mice were sacrificed, the cerebral infarct volume was measured by triphenyltetrazolium chloride staining. The brain water content was measured by dry and wet weight method. The expression of Nrf2 pathway related factors, including Nrf2, heme oxygenase 1 (HO-1) and NAD(P)H quinone oxidoreductase 1 (NQO1) were measured by quantitative real-time PCR and Western blotting. Results:On the 10 th day after modeling, compared with the Nrf2 -/- control group, the neurological deficit was significantly milder, the volume of cerebral infarction and brain water content were significantly smaller, and the mRNA and protein levels of Nrf2, HO-1 and NQO1 were significantly higher in the Nrf2 + /+ control group, and the differences were statistically significant ( P<0.05). For Nrf2 + /+ mice, compared with the control group, the cerebral infarct volume was significantly reduced ( P<0.05), the brain water content was significantly reduced ( P<0.05), and the neurological function recovery was significantly better ( P<0.05), and the levels of Nrf2, HO-1, and NQO1 mRNA and protein were significantly higher in the high-dose butylphthalide group (all P<0.05). For Nrf2 -/- mice, there were no significant differences in neurological function, cerebral infarction group volume, brain water content, Nrf2, HO-1, NQO1 mRNA and protein levels among the groups. Conclusion:Long-term butylphthalide pretreatment can significantly improve the neurological function, reduce cerebral infarction volume, reduce brain water content, and increase Nrf2, HO-1, NQO1 mRNA and protein expression levels in mice with permanent distal middle cerebral artery occlusion, suggesting butylphthalide may play a neuroprotective effect by up-regulating the expression of Nrf2 gene and its downstream antioxidant stress factors HO-1 and NQO1.

Objective: To reveal the related factors of inhibitors and differences ofhemorrhage and joint disease before and after the production of inhibitors in children with hemophilia A (HA) . Methods: Retrospective analyses of the clinical data of 381 children with HA under the age of 16 registered in the Registration Management Center of Hemophilia in Henan Provincial from January 2015 to August 2018. Results: A total of the 381 children were enrolled with 116 (30.4%) mild, 196 (51.4%) moderate, and 69 (18.1%) severe cases; 54 patients (14.2%) had inhibitors, including 22 high and 32 low titer inhibitors. Positive family history was positively associated with inhibitors[P<0.001, OR=3.299 (95%CI 1.743-5.983) ], and high-intensity exposure was associated with inhibitors[P=0.002, OR=2.587 (95%CI 1.414-4.731) ]. High-intensity exposure was associated with high titer inhibitor production[P=0.001, OR=8.689 (95%CI 2.464-30.638) ], and high-intensity exposure increased the risk of high titer inhibitors in HA patients. After inhibitors occurred in 54 patients with HA, the rates of overall joint annual bleeding (z=-3.440, P=0.001) and traumatic annual bleeding (z=-2.232, P=0.026) increased, but the rates of the annual joint bleeding (z=-1.342, P=0.180) and spontaneous annual bleeding (z=-1.414, P=0.157) remained to be not statistically significant. The joint ultrasound score did not change significantly after the inhibitor information (z=-0.632, P=0.527) . Conclusions Positive family history and high-intensity exposure could increase the risk of F Ⅷ inhibitors in HA patients, and high-intensity exposure increased the risk of high titer inhibitors. The rates of the overall joint annual bleeding and traumatic annual bleeding increased after the inhibitor information.

Objective To evaluate the role of 2B-containing NMDA receptors(NR2B)in sevoflu-rane anesthesia-induced cognitive dysfunction in aged rats.Methods Thirty-two healthy male Sprague-Dawley rats,aged 18 months,weighing 570-630 g,were divided into 4 groups(n＝8 each)using a ran-dom number table method: control group(group C),sevoflurane anesthesia group(group S),sevoflurane anesthesia plus NR2B specific inhibitor Ro 25-6981 group(group S+RO)and Ro 25-6981 group(group RO).S and S+RO groups inhaled 3％sevoflurane for 4 h.Ro 25-6981 1 mg/kg was intraperitoneally injec-ted at 15 min before inhaling sevoflurane in group S+RO.Morris water maze test was performed at 2 days af-ter the end of anesthesia to assess cognitive function.The rats were then sacrificed,and hippocampal tis-sues were obtained for determination of the expression and phosphorylation of ERK1/2 by Western blot.Results Compared with group C,the escape latency was significantly prolonged,the frequency of crossing the original platform was reduced,the time of staying at the original platform quadrant was shortened,and the phosphorylation of ERK1/2 was decreased in group S(P＜0.05),and no significant change was found in the escape latency in S+RO and RO groups(P>0.05).Compared with group S,the escape latency was significantly shortened,the frequency of crossing the original platform was increased,the time of staying at the original platform quadrant was prolonged,and the phosphorylation of ERK1/2 was increased in group S+RO(P＜0.05).There was no significant difference in ERK1/2 expression among the four groups(P>0.05).Conclusion The mechanism by which sevoflurane anesthesia induces cognitive dysfunction is re-lated to up-regulating the expression of NR2B and inhibiting the activity of ERK1/2 in aged rats.

Objective: To analyze the percentage of myeloperoxidase (MPO)-positive acute myeloid leukemia (AML) blast cells, and to explore the correlation of MPO expression with the clinical features, gene alterations, therapeutic response and prognosis of AML. Methods: The expressions of MPO in BM blasts cells of 233 newly diagnosed AML were retrospectived analyzed, they were divided into two groups using the percentage of MPO-positive blast [low (≤70%) and high (>70%)], clinical features, gene alterations, chemotherapy efficacy and prognosis were compared between the two groups. Results: ①Of the 233 patients, 121(51.9%) were in the low MPO group, and the rest 112(48.1%) in the high MPO group. Favorable-risk group according NCCN guidelines of AML was always MPO-high (χ²=32.773, P<0.001), while MPO-low was closely related to poor-risk (χ²=7.078, P=0.008); ②DNMT3A mutation (χ²=6.905, P=0.009), spliceosome genes mutation (SF3B1/SRSF2/U2AF1) (χ²=5.246, P=0.022), RUNX1 mutation (χ²=4.577, P=0.032), ASXL1 mutation (χ²=7.951, P=0.005) and TP53 mutation (P=0.004) were more likely to be seen in the low MPO group, while C-KIT mutation (χ²=8.936, P=0.003) and CEBPA mutation (χ²=12.340, P<0.001) were more frequent in the high MPO group, especially CEBPA double mutation; ③The rates of first complete remission in the low MPO group were significantly lower than that in the high MPO group (38.8% vs 68.1%, χ²=15.197, P<0.001). Multivariate analysis showed that low MPO positivity significantly affected the CR₁ unfavourably. ④The overall survival (OS) and the progression-free survival (PFS) were significantly worse in the low MPO group (18.0% vs 89.4% for OS, and 11.5% vs 56.7% for PFS, P<0.001). Multivariate analysis disclosed that the low number of MPO was significantly unfavourable prognostic factor. ⑤The low MPO group still showed a worse survival even when restricted to the patients with normal karyotype, the OS and the PFS were 31.1% and 18.8% respectively. Conclusions AML with different MPO expression percentage had a unique gene mutation spectrum. Low expression of MPO was an independent risk factor for CR₁, OS and PFS in AML patients, which may be a simple and highly significant factor for AML patients when evaluating the therapeutic efficacy and prognosis.

Objective: To evaluate the role of 2B-containing NMDA receptors (NR2B) in sevoflurane anesthesia-induced cognitive dysfunction in aged rats. Methods: Thirty-two healthy male Sprague-Dawley rats, aged 18 months, weighing 570-630 g, were divided into 4 groups (n=8 each) using a random number table method: control group (group C), sevoflurane anesthesia group (group S), sevoflurane anesthesia plus NR2B specific inhibitor Ro 25-6981 group (group S+ RO) and Ro 25-6981 group (group RO). S and S+ RO groups inhaled 3% sevoflurane for 4 h. Ro 25-6981 1 mg/kg was intraperitoneally injected at 15 min before inhaling sevoflurane in group S+ RO.Morris water maze test was performed at 2 days after the end of anesthesia to assess cognitive function.The rats were then sacrificed, and hippocampal tissues were obtained for determination of the expression and phosphorylation of ERK1/2 by Western blot. Results: Compared with group C, the escape latency was significantly prolonged, the frequency of crossing the original platform was reduced, the time of staying at the original platform quadrant was shortened, and the phosphorylation of ERK1/2 was decreased in group S (P<0.05), and no significant change was found in the escape latency in S+ RO and RO groups (P>0.05). Compared with group S, the escape latency was significantly shortened, the frequency of crossing the original platform was increased, the time of staying at the original platform quadrant was prolonged, and the phosphorylation of ERK1/2 was increased in group S+ RO(P<0.05). There was no significant difference in ERK1/2 expression among the four groups (P>0.05). Conclusion The mechanism by which sevoflurane anesthesia induces cognitive dysfunction is related to up-regulating the expression of NR2B and inhibiting the activity of ERK1/2 in aged rats.

Objective To determine collateral circulation in patients with acute ischemic stroke using capillary index score (CIS)in order to evaluate the prognosis of endovascular treatment. Methods From January 2013 to December 2015,46 consecutive patients with acute ischemic stroke treated with endovascular treatment at the Department of Neurology,Central Hospital of Baotou were enrolled retrospectively. Angiography was performed before endovascular treatment in order to complete CIS score. The patients were divided into a good prognosis group (n = 21)and a poor prognosis group (n = 25)according to the modified Rankin scale (mRS)scores. Univariate analysis was used to compare the baseline data and the clinical data of the two groups,including age,sex,history of diabetes,pretreatment systolic blood pressure,conducting intravenous thrombolysis or not,time from ictus to intravenous thrombolysis,National Institutes of Health Stroke Scale (NIHSS)score,Alberta stroke program early CT score (ASPECTS),vascular filling,time from onset to revascularization,and postoperative vascular recanalization (the modified Thrombolysis in Cerebral Infarction [mTICI]). Multivariate analysis was used to analyze the effect of CIS score on good prognosis. Results There were no significant differences in age,sex,history of diabetes,pretreatment systolic blood pressure,conducting intravenous thrombolysis or not,time from ictus to thrombolysis,and number of mechanical thrombectomy between the good prognosis group and the poor prognosis group (all P > 0. 05). There were significant differences in the NIHSS score (15 ± 3 vs. 19 ± 4),ASPECTS score (8 [7,10]vs. 6 [5,8]),filling well 85. 7% (18 / 21)vs. 44. 0% [11 / 25]),time from ictus to recanalization (363 ± 42 min vs. 398 ± 53 min),and postoperative vascular recanalization (mTICI≥Ⅱb)(100. 0% [21 / 21]vs. 68. 0%[17 / 25];all P < 0. 05). CIS (OR,8. 600,95% CI 2. 670 -33. 800)and mTICI grade (OR,5. 720, 95%CI 12. 170-22. 300)were significantly associated with the prognosis. Conclusion The CIS score can be used to evaluate brain perfusion. fCIS is closely associated with the good clinical prognosis. When screening the suitable patients for endovascular therapy,increasing the CIS score to evaluate the salvageable brain tissue is effective and feasible.

Objective To investigate the effect of high frequency (10 Hz),low frequency (1 Hz) and theta burst stimulation (TBS) mode of repetitive transcranial magnetic stimulation (rTMS) on the recovery of motor function in hemiplegic patients following acute ischemic stroke.Methods Seventy-two patients with hemiplegia after acute ischemic stroke were randomly grouped with the random number table.They were treated with low frequency (n=18),high frequency (n=18),and TBS (n=18) rTMS or sham stimulation (control group,n=18),once a day,for 2 weeks.Fugl-Meyer Assessment (FMA) and National Institutes of Health Stroke Scale (NIHSS) were used to evaluate neurological function in all patients before rTMS treatment (on the day before the first treatment) and after treatment (on the day after the last treatment).Results After treatment,the FMA and NIHSS scores in the 4 groups were significantly improved compared with before treatment (all P<0.05).After rTMS treatment,the FMA and NIHSS scores were improved significantly in the high frequency group,low frequency group and TBS group compare with the control group (all P<0.05).There were no significant differences among all the treatment groups.Conclusion sHigh frequency,low frequency and TBS rTMS can improve the recovery of motor function in hemiplegic patients following acute ischemic stroke.There were no significant differences among all the treatment modes.

Objective To investigate the clinical features,etiology,diagnosis and treatment of acute auditory agnosia.Methods We studied the clinical manifestation,diagnosis and treatment of acute auditory agnosia in a patient in our hospital.Results A 28 year oldyoung woman visited our department because she suffered from the tinnitus for 7 days and she could not distinguish the semantics for 1 day.There were no other abnormal symptoms in the central and peripheral nervous system on admission.Audiological testing showed normal,language testing showed that the speech discrimination score was zero.MRI showed extensive damage to temporal lope.MR spectroscopy revealed increased lactate and reduced N-acetyl aspartate.Acute auditory agnosia resulted from mitochondrial myopathy was considered.After symptomatic treatment,the symptoms were significantly improved.Molecular genetics examination showed the A3243G mtDNA mutation,further confirmed the diagnosis of mitochondrial encephalomyopathy with lactic acidosis and stroke like episodes (MELAS) syndrome.Conclusion Acute auditory agnosia and acute tinnitus can be the first symptoms in MELAS,thus,MELAS should be suspected in patients with acute auditory agnosia,acute tinnitus,sudden hearing loss in children and youth.Imaging examination plays an important role in the etiological diagnosis of acute auditory agnosia.