Objective To investigate the effect of hypoxic preconditioning （HPC） on human umbilical vein endothelial cells（HUVECs） at the cellular level （cell morphology， viability， and apoptosis rate）， the molecular level [the expression of vascular endothelial growth factor（VEGF）and angiogenin-2（Ang-2）]， and the genetic level （the expression of corresponding mRNAs）， and to discuss the protective mechanism of HPC against hypoxic injury of HUVECs. Methods HUVECs were routinely cultured to establish a hypoxia model of HUVECs， and then HUVECs were divided into normal control group （CON group），hypoxia group （HYP group）， and HPC group. Cell morphology was observed for each group under a microscope，and CCK-8 assay， Hoechst33258 fluorescent staining， Western blotting， and RT-PCR were used to measure cell viability， cell apoptosis rate， and the expression levels of VEGF， Ang-2， and corresponding mRNAs in cells. Results Compared with the CON group， the HYP group had significant damage of cells， a significant reduction in cell viability， and significant increases in cell apoptosis rate and the levels of VEGF， Ang-2，and corresponding mRNAs in cells， while the HPC group had basically normal cell morphology， significantly lower cell viability and apoptosis rate than the CON group， and significantly higher cell viability and apoptosis rate than the HYP group， as well as significantly higher expression levels of VEGF， Ang-2，and corresponding mRNAs than the CON group and the HYP group. Conclusion HPC exerts a protective effect against hypoxic injury of HUVECs by upregulating the expression of VEGF，Ang-2，and corresponding mRNAs.

Objective To investigate the level of inflammation after interventional treatment in patients undergoing intracranial stent implantation by measuring the changes in the plasma levels of monocytes and high-density lipoprotein cholesterol（HDL-C） and monocyte to high-density lipoprotein cholesterol ratio （MHR） after stent implantation for intracranial atherosclerotic stenosis （ICAS） in high-altitude areas， as well as the causes of such changes and their value in predicting clinical prognosis. Methods The ICAS patients who were consecutively admitted to Qinghai Provincial People’s Hospital， from June 10， 2017 to March 1， 2022 and underwent interventional treatment were enrolled， and all patients signed the informed consent. Clinical data and the data on interventional surgery were collected， and blood samples were collected before interventional treatment， within 72 hours after interventional treatment， and at 3 months after interventional treatment to measure the levels of monocytes and HDL-C. The above indicators were compared before and after interventional treatment， and such changes were analyzed in terms of their association with the site of cerebrovascular stenosis and NIHSS score. Results A total of 123 patients with severe ICAS who underwent intracranial stent implantation and had complete data were included. Compared with the data before surgery， there was a significant increase in the plasma level of monocytes at 72 hours after stent implantation [（0.64±0.21）×109/L vs （0.53±0.17）×109/L， P<0.001]， while there was a significant reduction in the plasma level of monocytes at 3 months after stent implantation [（0.43±0.14）×109/L vs （0.53±0.17）×109/L，P<0.001]. Compared with the data before surgery， there was no significant change in HDL-C within 72 hours after surgery[（0.93±0.21）mmol/L vs （0.93±0.18）mmol/L， P>0.005]， while there was a significantly increase in HDL-C at 3 months after surgery[（1.05±0.21 mmol/L vs （0.93±0.18）mmol/L， P<0.001）. There was no significant correlation between monocytes/HDL-C/MHR and NIHSS score before surgery and at 72 hours after surgery （P>0.005）； there was no significant correlation between monocytes/HDL-C/MHR and NIHSS score before surgery and within 72 hours after surgery （P>0.005）；at 3 months after surgery， monocytes and MHR were significantly negatively correlated with NIHSS score （r=-0.271，P<0.05；r=-0.320，P<0.005），while HDL-C was significantly positively correlated with NIHSS score （r=0.213， P<0.001）. Conclusion Balloon dilatation and ischemia/reperfusion after intracranial stent implantation may cause inflammatory response in the body， thereby leading to increases in the level of monocytes and the value of MHR. Therefore， monocytes， HDL-C， and MHR may be used as predictive factors for the improvement in neurological defects in the convalescence stage after stent implantation.
Monocytes

Objective:To investigate the distribution of intraocular pressure (IOP) in high-altitude population aged 18 years and over in Xining, Qinghai and establish the reference interval (RI) of IOP.Methods:A cross-sectional study was conducted in Xining, Qinghai Province at 2.271 km above sea level from September 2019 to May 2020.Ophthalmic examinations and IOP measurement were conducted among subjects from Physical Examination Center of Qinghai Provincial People's Hospital.The subjects who had been living in Xining without leaving for three months were enrolled.Ophthalmic examinations included vision examination, IOP measurement, slit-lamp microscopy, fundus photography, anterior and posterior segment optical coherence tomography.IOP was measured using Goldmann applanation tonometry under local anesthesia.Subjects with factors that could cause significant changes in IOP and affect the accuracy of IOP measurement, and those who were unable to receive IOP measurement were excluded.Subjects were grouped according to sex, age and ethnicity, and the distribution and RI of IOP were compared among all groups.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Beijing Tongren Hospital, Capital Medical University (No.TRECKY2017-024). Written informed consent was obtained from each subject.Results:A total of 6 120 subjects (6 120 eyes) aged 18-90 years old were enrolled, including 2 850 males and 3 270 females with average age of (45.54±13.85) years.The average IOP of high-altitude population in Xining, Qinghai Province was (14.32±1.93) mmHg (1 mmHg=0.133 kPa), with the RI of 10.54-18.10 mmHg.The average IOP was (14.42±1.98) mmHg in male with the RI of 10.54-18.30 mmHg, (14.23±1.88) mmHg in female with the RI of 10.55-17.91 mmHg.The IOP of male was higher than that of female ( t=3.71, P<0.001). The IOP of Han, Tibetan, Hui and other nationalities were (14.38±1.91), (13.93±2.06), (14.21±1.87), (13.94±1.95) mmHg, respectively, with a statistically significant overall difference ( F=6.73, P<0.001). The IOP of Han nationality was significantly higher than that of Tibetan, Hui and other nationalities, and the differences were statistically significant (all at P<0.05). Conclusions:RI of IOP in high-altitude population from Xining, Qinghai is lower compared with normal altitude area.

Objective To investigate the MRI manifestations and analyze the prognostic factors of patients with anterior circulation minor stroke and nonGminor stroke in Qinghai plateau.Methods 41 6 cases of the first admission,including 1 9 2 patients with minor stroke and 224 patients with nonGminor stroke.MRI and MRA examinations of the head were completed in all patients within 72 h of admission.Patients were followed up for one year to observe the recurrence of stroke,and the quality of life was evaluated with the help of modified Rankin Scale (MRS)scores.Results (1)MRA showed that 36.98% of the minor stroke and 58.93% of the nonGminor stroke had the stenosis of the responsible artery at the infarction site.The difference was significant (χ2＝ 1 9.94,P＜ 0.00 1 ).(2 )MRI showed that the initial infarction sites of minor stroke and nonGminor stroke were different (χ2＝4.47 ,P＜0.005 ).(3 )The recurrence rate was 10.42% in minor stroke and 12.05% in nonGminor stroke.There was no significance between the two groups (χ2＝ 0.28,P＞0.05).(4) Among patients with poor prognostic outcomes (whose MRS≥3),there were 1 9 cases of minor stroke and 6 1 cases of nonGminor stroke,and the difference was significant (χ2＝20.00,P＜0.0 1 ).Conclusion LesionGrelated vascular stenosis in patients with minor stroke is mild and the primary infarction is more common in isolated subcortical or deep white matter.The stenosis is severe in patients with nonGminor stroke,and the infarct lesion is often subcortical with or without cortical or deep white matter.There is no difference in recurrence risk between minor stroke and nonGminor stroke.The prognosis of minor stroke is better than that of nonGminor stroke.

Objective To discuss the correlation of apolipoprotein E (ApoE) gene polymorphism with cerebral infarction (CI)and intracerebral hemorrhage (ICH) among Tibetan nationality with cerebrovascular diseases in Qinghai Province, seek the differences in each allele of ApoE in Tibetan nationality. Methods The data from a total of 94 patients with cerebrovascular diseases was collected from the people's hospital of qinghai province, the people's hospital of guoluo prefecture , and the people's hospital of yushu prefecture as the cerebrovascular disease group, including 48 cases of cerebral infarction. There were 46 cases of cerebral hemorrhage. A total of 96 healthy Tibetan subjects were selected as the control group. DNA was extracted from all subjects. Real－time PCR was used to detect ApoE. The correlation between ApoE genotype and cerebral infarction and cerebral hemorrhage was analyzed. Results E2/E3 gene was common in Tibetan nationality with cerebrovascular diseases. E2/E3 genotype accounted for 50% in cerebral infarction group. E2/E3 (65.2%) was the most common in intracerebral hemorrhage group. E2/E4 (64.6%) was the most common in the control group. There was a statistically significant difference between the two groups (P＜0.01). In the Tibetan population, ε3 allele genome (48.0%) was the most common in cerebral infarction group and ε2(43.5%) were the most common alleles in intracerebral hemorrhage group. In the normal control group, ε4 (49.0%) was the most common allele. Conclusion E2/E3 genotype may be related to cerebrovascular diseases. ε3 allele may be the susceptible factor of cerebral infarction wherase ε4 may be the protective factor of cerebrovascular diseases in Tibetan population.

Objective To investigate the effect of seasonal and meteorological factors on the onset of stroke in Qinghai Plateau area.Methods Patients with stroke admitted to Qinghai Provincial People's Hospital from December 1,2011 to November 30,2016 were enrolled retrospectively.The meteorological data provided by Qinghai Meteorological Bureau were used to analyze the distribution of the number of cases in different seasons in Qinghai area and the relationship between the incidence of stroke and plateau meteorological factors.Results A total of 9 412 patients with acute ischemic stroke were enrolled,aged 15～95 years.There were significant statistical differences in the number of cases of ischemic stroke,cerebral hemorrhage,and subarachnoid hemorrhage in different seasons (all P ＜ 0.05).Among them,the number of cases with ischemic stroke increased first and then gradually decreased with the changes of spring,summer,fall and winter,while cerebral hemorrhage and subarachnoid hemorrhage showed a trend of decreasing first and then increasing gradually.There were significant differences in the number of cases in different relative humidity,temperature,and temperature differences (all P ＜ 0.05).Conclusion There are seasonal differences in the incidence of stroke in the plateau area,which may be associated with the influence of plateau meteorological factors.

Migraine is a common neurological disease.It involves complex neurological abnormalities.Recent advances in the neurophysiology of migraine have enabled us to explain some of the symptomatic problems and have contributed to the development of new targeted treatments that may change the way migraine treated in the future.Migraine treatment is individualized,in which preventive drug therapy also plays an important role.This article will discuss the new progress in the treatment of migraine,with emphasis on the new treatment of calcitonin gene-related peptide pathway.

Our previous works disclosed the contributing role of macrophage migration inhibitory factor (MIF) and dopaminergic inhibition by lysine dimethyltransferase G9a/Glp complex in peripheral nerve injury-induced hypersensitivity. We herein propose that the proinflammatory cytokine MIF participates in the regulation of neuropathic hypersensitivity by interacting with and suppressing the descending dopaminergic system. The lumbar spinal cord (L-SC) and ventral tegmental area (VTA) are two major locations with significant upregulation of MIF after chronic constriction injury (CCI) of the sciatic nerve, and they display time-dependent changes, along with a behavioral trajectory. Correspondingly, dopamine (DA) content shows the reverse characteristic change to MIF with a time-dependent curve in post-surgical behavior. The levels of both MIF and DA are reversed by the MIF tautomerase inhibitor ISO-1, and a negative relationship exists between MIF and DA. The reversed role of ISO-1 also affects tyrosine hydroxylase expression. Furthermore, CCI induces Th promoter CpG site methylation in the L-SC and VTA areas, and this effect could be abated by ISO-1 administration. G9a/SUV39H1 and H3K9me2/H3K9me3 enrichment within the Th promoter region following CCI in the L-SC and VTA was also decreased by ISO-1. In cultured dopaminergic neurons, rMIF enhanced the recruitment of G9a and SUV39H1, followed by an increase in H3K9me2/H3K9me3. These molecular changes correspondingly exhibited alterations in Th promoter CpG site methylation and pain behaviors. In summary, MIF functions as a braking factor in curbing dopaminergic descending inhibition in peripheral nerve injury-induced hypersensitivity by mediating Th gene methylation through G9a/SUV39H1-associated H3K9 methylation.

Objective To investigate the roles of phosphorylated glycogen synthase kinase 3β (pGSK3β) and phosphorylated signal transducer and activator of transcription 3 (pSTAT3) in hypoxic preconditioning-induced neuroprotection against ischemic brain injury in rats.Methods Sixty SD rats were randomly divided into a sham operation group,a cerebral ischemia group,and a hypoxia preconditioning group (n =20 in each group).A model of middle cerebral artery occlusion (MCAO) was induced by the modified suture method.Before the preparation of MCAO model,the rats in the hypoxia preconditioning group were put into a hypobaric oxygen chamber at a simulated altitude of 5 000 m (pressure:0.53 × 105 kPa;partial pressure of oxygen:81 mmHg;1 mmHg =0.133 kPa),3 h a day for 5 days.At 24 h after MCAO modeling,the rats were subjected to neurobehavioral score (n =6) and cerebral infarction volume measurement (n =6).Immunohistochemical staining was used to detect the expression levels of neuronal nuclei (NeuN) and pGSK3β (Ser9) (n=7).Western blot was used to detect the expression levels of pGSK3 β (Ser9) and pSTAT3 (Tyr705) in the ischemic cortex (n =7).Results The neurological deficit score (1.833 ±0.408 vs.2.667 ± 0.516;t =3.101,P=0.011) and cerebral infarction volume (18.137％ ± 0.801％ vs.24.125％ ± 0.694％;t =13.840,P＜ 0.001) in the hypoxia preconditioning group were significantly lower or smaller than those in the cerebral ischemia group.Immunohistochemical staining showed that the numbers of NeuN positive cells in the cerebral ischemia group and the hypoxia preconditioning group were significantly less than that in the sham operation group (48.000 ± 1.414/high power field [HPF],124.833 ± 3.061/HPF,and 213.500 ± 2.429/HPF;F =7 150.550,P ＜ 0.001),the hypoxia preconditioning group was significantly more than the ischemia group (P ＜0.001);the numbers of pSTAT3 positive cells in the cerebral ischemia group and the hypoxia preconditioning group were significantly higher than that in the sham operation group (57.667 ± 1.366/HPF,29.167 ± 1.941/HPF and 3.500 ± 1.049/HPF;F =1 962.649,P ＜0.001),and the hypoxia preconditioning group was significantly less than the ischemia group (P ＜0.001).Western blot analysis showed that the expression levels of ischemic cortical pGSK3β and pSTAT3 in the cerebral ischemia group and the hypoxia preconditioning group were significantly higher than those of the sham operation group (pGSK3 β:2.336 ± 0.102,0.876 ± 0.196 and 0.440 ± 0.012;F =1 610.826,P ＜ 0.001;pSTAT3:8.368± 0.230,4.883± 0.123 and 0.595± 0.138;F=4018.051,P＜0.001),the hypoxia preconditioning group were significantly lower than the ischemia group (all P ＜0.001).Conclusions Hypoxia preconditioning has neuroprotective effect for ischemic brain injury in rats.It may be associated with the down-regulation of the expressions of pGSK3 and pSTAT3.

Objective To investigate the influence of the chronic mountain sickness (chronic mountain sickness, CMS) on Cerebrovascular reactivity (CVR) and regulators of cerebrovascular responses. Methods Twenty-six CMS pa?tients and 23 healthy control group using transcranial Doppler ultrasound (Transcranial Doppler Ultrasoiund, TCD) as?sessment of CVR, enzyme-linked immunosorbent assay were applied to measure the serum levels of the endothelin (endo?thelin 1 , ET-1) and its receptor (endothelin receptor B, ETBR), endothelial nitric oxide synthase (endothelial nitric oxide synthase, eNOS) in CMS patients and healthy control. Results CVR (3.84 ± 3.01) was significantly lower in CMS patients than in the healthy control CVR (6.39 ± 6.87) (P <0.05); the serum concentration of ETBR in CMS patients was higher than in control [(386.07±281.57)μg/μL vs.(312.30±238.07)μg/μL] (P<0.05). Conclusions The cerebral circulation re?serve in CMS patients is significantly lower compared with healthy people. The regulation of vascular capacity by serum ET-1 and eNOS is similar between CMS patients and healthy control. The hypoxic vasodilatation in CMS patients is close?ly associated with cerebrovascular high expression of ETBR. This study may provide a scientific basis for the prevention and treatment of cerebral infarction in the patients with chronic mountain sicknes.