Aim To study the proliferative effect of resveratrol pretreatment on oxygen-glucose deprivation/reoxygenation ( OGD/R ) injury of rat cortical neural stem cells ( NSCs ) in vitro. Methods Isolation and purification of NSCs in neonatal Sprague-Dawley( SD) rats were conducted by suspended cultivation. The third passage NSCs of adherent culture was cultured under oxygen and glucose deprivation for 150 min and reoxygenation for 24 h. The experimental subjects were divided into normal, control, ethanol and resveratrol pretreatment groups. Immunofluorescence was used to identify NSCs. Cell viability was detected with CCK-8 assay. Flow cytometry cell cycle and BrdU assay were used to measure cell proliferation. Results Cells both in suspended and adherent cultivation highly expressed neuroepithelial stem cell protein ( nestin ) . Compared with the control group, NSCs viabilities and prolifera-tion in resveratrol groups (1, 5, 20 μmol·L-1 ) were significantly heightened, and highest in the 5 μmol · L-1 resveratrol group ( P<0. 05 ) . Conclusion Res-veratrol pretreatment can reduce injury and promote proliferation of NSCs after oxygen-glucose deprivation /reoxygenation.

The M and S segment cDNAs of hantavirus Z5 strain was amplified by RT-PCR,and the purified PCR products were cloned into vector pGEM-T and then sequenced.It was demonstrated that the M genome segment of Z5 was found to be 3 616 nucleotides in length with a single open reading frame encoding 1 135 amino acids.And the S genome segment was 1700 nucleotides in length with a single open reading frame encoding 429 amino acids.As demonstrated by the homologous analysis of nucleotides and amino acids,it was showed that the Z5 strain belonged to hantaan viruses HTN type and was the same subtype of the Z10 strain.It is conclouded that difference in nucleotide sequence exists between Z5 strain with other Hantavirus strains but high level of homology in amino acid sequences is still present.

Objective:To investigate the expressions and clinical significance of tetraspanin CO-029 and integrin αv in intrahepatic cholangiocarcinoma (ICC ).Methods:Tissue microarray (TMA) was used to detect the expression of CO-029 and αv in 254 cases of intrahepatic cholangiocarcinoma. The relationship between the two factors and clinicopathological features, recurrence, metastasis and prognosis was analyzed.Spearman method was used to analyze their correlation.Relationship between αv and CO-029 was studied by mass spectrometry and database search,immunoprecipitation and Western blot were used to detect the coexistence.Results:Tissue microarray analysis showed that the positive expression rate of CO-029 was 51.6% (131/254), and the positive expression rate of αv was 61.4% (156/254). The expression of CO-029 and αv were closely correlated with tumor envelope, size, number and TNM stage ( P<0.05). According to the time of recurrence (TTR), the expressions of CO-029 and αv in early postoperative recurrence group (TTR <1 year) were significantly higher than those in non recurrence group (TTR ≥ 1 year). The patients with high CO-029 expression were more likely to relapse ( HR=2.01, 95% CI=1.45-2.79; P<0.001) and had shorter survival time ( HR=2.03, 95% CI=1.46-2.81; P<0.001). The patients with high expression of αv had shorter recurrence time ( HR=1.85, 95% CI=1.38-2.47; P<0.001) and shorter survival time ( HR=1.95, 95% CI=1.40-2.71; P<0.001). Co immunoprecipitation and Western blot confirmed that αv and CO-029 formed a complex. There was a positive correlation between CO-029 and αv in intrahepatic cholangiocarcinoma ( r=0.401, P<0.01). Conclusions:The differential expression of CO-029 and αv were closely related to the recurrence, metastasis and prognosis of intrahepatic cholangiocarcinoma, and CO-029 may couple with αv to form a complex to promote the invasion and metastasis of intrahepatic cholangiocarcinoma.

ObjectiveTo investigate the effect of α-Zearalanol(α-ZAL) on lipometabolism and hemorheology in ovariectomized(OVX) hyperlipidemia rabbits.Methods44 adult virgin female rabbits were divided into 5 groups,group A: normal control;group B: sham+CHO;group C: OVX+CHO;group D: OVX+CHO+17βE_2;group E: OVX+CHO+α-ZAL.Cholesterol(CHO) was fed to rabbits for 12 weeks.Before and after feeding CHO,the serum lipid(TC,TG,LDL-C,HDL-C) were measured;Blood viscosity,plasma viscosity,aggregation index of RBC(AIRC) and fibrinogen were also assayed respectively.ResultsThe serum levels of TC,TG and LDL-C in group B and E were significantly decreased compared with those in group C(P<0.05);the level of blood viscosity,plasma viscosity and AIRC platelet aggregation rate in group D and E were also significantly decreased compared with those in group C(P<0.05).Conclusionα-ZAL can improve vascular function through the adjustment of lipometabolism and hemorheology.

Objective To investigate the status quo of the cerebral injury in patients with acute stress disorder (ASD) and the impact of family function,mental toughness,and the characteristics of brain lesions on it.Methods A total of 349 patients were enrolled from North China University of Science and Technology affiliated hospital neurology department and neurosurgery department from May 2016 to November 2016 and they were tested with Stanford Acute Stress Reaction Questionnaire (SASRQ),the family function assessment scale (APGAR),Chinese version of mental toughness scale (CD-RISC).Results 349 cases of patients with SASRQ score (57.21±44.97),8 to 39 160 people (45.8％),40 to 56 85 people (24.4％),57 to 150 104 people (29.8％).The results showed that whether the hemiplegia (β=-0.030),family function (β=-0.032),mental toughness (β=0.886),disturbing degree (β=0.052),bad days (β=0.060)were picked in the regression equation (P＜0.05).Conclusion There were obvious acute stress symptoms in patients with brain injury.The individuals who have family dysfunction,the worse the psychological resilience and the higher the disturbance degree of the patients with hemiplegia,may be likely to get acute stress disorder.

Background: Coinfection with avian leukosis virus subgroup J (ALV-J) and reticuloendotheliosis virus (REV) is common in chickens, and the molecular mechanism of the synergistic pathogenic effects of the coinfection is not clear. Exosomes have been identified as new players in the pathogenesis of retroviruses. The different functions of exosomes depend on their cargo components. Objectives: The aim of this study was to investigate the function of co-regulation differentially expressed proteins in exosomes on coinfection of ALV-J and REV. Methods: Here, viral replication in CEF cells infected with ALV-J, REV or both was detected by immunofluorescence microscopy. Then, we analyzed the exosomes isolated from supernatants of chicken embryo fibroblast (CEF) cells single infected and coinfected with ALV-J and REV by mass spectrometry. KEGG pathway enrichment analyzed the co-regulation differentially expressed proteins in exosomes. Next, we silenced and overexpressed tripartite motif containing 62 (TRIM62) to evaluate the effects of TRIM62 on viral replication and the expression levels of NCK-association proteins 1 (NCKAP1) and actin-related 2/3 complex subunit 5 (ARPC5) determined by quantitative reverse transcription polymerase chain reaction. Results: The results showed that coinfection of ALV-J and REV promoted the replication of each other. Thirty proteins, including TRIM62, NCK-association proteins 1 (NCKAP1, also known as Nap125), and Arp2/3-5, ARPC5, were identified. NCKAP1 and ARPC5 were involved in the actin cytoskeleton pathway. TRIM62 negatively regulated viral replication and that the inhibition of REV was more significant than that on ALV-J in CEF cells coinfected with TRIM62. In addition, TRIM62 decreased the expression of NCKAP1 and increased the expression of ARPC5 in coinfected CEF cells. Conclusions Collectively, our results indicated that coinfection with ALV-J and REV competitively promoted each other's replication, the actin cytoskeleton played an important role in the coinfection mechanism, and TRIM62 regulated the actin cytoskeleton.

Objective:To evaluate the role of Rac1 in cerebral ischemia-reperfusion (I/R) injury and the relationship with mitopaghy in diabetic rats.Methods:SPF healthy adult male Sprague-Dawley rats, aged 8 weeks, weighing 250-280 g, in which diabetes mellitus was induced by intraperitoneal streptozotocin, were used in this study.Forty-eight rats with diabetes mellitus were divided into 4 groups ( n=12 each) using a random number table method: sham operation group (sham group), cerebral I/R group (I/R group), I/R plus lentivirus inhibiting Rac1 group (I/R+ shRac1 group), and I/R plus lentivirus-negative control group (I/R+ NC group). Cerebral I/R was induced by 90-min middle cerebral artery occlusion followed by 24-h reperfusion.In I/R+ shRac1 and I/R+ NC groups, Rac1 shRNA lentivirus vector and lentivirus negative control vector 10 μl were injected via the right lateral cerebral ventricle at 7 days before establishing the model, respectively.Rats were sacrificed at 24 h of reperfusion, and brains were removed for determination of cerebral infarct size, expression of BNIP3, P62, LC3Ⅰ and LC3Ⅱ (by Western blot). The LC3Ⅱ/LC3 Ⅰ ratio was calculated. Results:Compared with sham group, the cerebral infarct size was significantly increased in the other three groups ( P<0.05). Compared with I/R group, the cerebral infarct size was significantly decreased, LC3Ⅱ/LC3Ⅰratio was increased, the expression of BNIP3 was up-regulated, and the expression of P62 was down-regulated in group I/R+ shRac1 ( P<0.05 or 0.01), and no significant change was found in each index in group I/R+ NC ( P>0.05). Compared with I/R+ NC group, the cerebral infarct size was significantly decreased, LC3Ⅱ/LC3Ⅰratio was increased, the expression of BNIP3 was up-regulated, and the expression of P62 was down-regulated in group I/R+ shRac1 ( P<0.05 or 0.01). Conclusion:The mechanism by which Rac1 reduces cerebral I/R injury is related to enhancing mitophagy in diabetic rats.

Objective:To explore the clinical characteristics and genetics of a Chinese patient with Gitelman syndrome (GS) and improve the awareness and diagnosis of GS among clinicians.Methods:Retrospectively analyzed the GS patient's clinical feature, laboratory examination, diagnosis, treatment and literature review admitted to Hebei General Hospital in September 2022.Results:A twelve-year-old boy was admitted to our department due to weakness of lower limbs. Laboratory tests after admission showed hypokalemia, hypomagnesemia, hypocalcemia and metabolic alkalosis. Genetic testing showed tow compound heterozygous mutations in the SLC12A3 gene (c.1456G>A and c.634G>A), which ultimately diagnosed as GS. The patient is on the mend and allowed to leave the hospital after treated by potassium supplement.Conclusion:The rate of leak diagnosis is high. Genetic testing should be undergo earlier if the patients suspected GS.

Objective To evaluate the role of long non-coding RNA maternally expressed gene 3(MEG3)in hyperglycose-induced neurocyte damage and the relationship with mitochondrion-dependent ap-optosis in rats.Methods Normally cultured PC12 cells were divided into 5 groups(n＝18 each)using a random number table method: normal concentration of glucose control group(C group),normal concentra-tion of glucose plus MEG3 group(C+MEG3 group),high-concentration glucose group(HG group),high-concentration glucose plus MEG3 group(HG+MEG3 group),and high-concentration glucose plus negative lentiviral vector(LV-NC)group(HG+NC group).PC12 cells were cultured in DMEM medium with 25 mmol/L glucose in group C.PC12 cells were cultured in DMEM medium with 25 mmol/L glucose after being transfected with MEG3 lentiviral vector(LV-MEG3)in C+MEG3 group.PC12 cells were cultured in DMEM medium with 250 mmol/L glucose in HG group.PC12 cells were incubated in DMEM medium con-taining 250 mmol/L glucose after being transfected with LV-MEG3 in HG+MEG3 group.PC12 cells were in-cubated in DMEM medium containing 250 mmol/L glucose after being transfected with LV-NC in HG+NC group.After the cells were cultured or incubated for 1 day,the cell viability was measured by CCK8 assay,the apoptosis rate and level of reactive oxygen species(ROS)were determined by flow cytometry,and the amount of lactic dehydrogenase(LDH)released was measured by DCFH-DA,the expression of Cyt c,caspase-3,caspase-9,Bcl-2,Bax and Apaf-1 was determined by Western blot,and the opening of mito-chondrial permeability transition pore(mPTP)was determined by fluorescent method.Blc-2/Bax ratio was calculated.Results Compared with group C,the cell viability was significantly decreased,the amount of LDH released,ROS level and apoptosis rate were increased,the opening of mPTP was increased,and the expression of caspase-3,caspase-9,Cyt c,Bax,Bcl-2 and Apaf-1 was up-regulated in HG,HG+MEG3 and HG+NC groups,and Bcl-2/Bax ratio was increased in HG+MEG3 group and decreased in HG and HG+NC groups(P＜0.05).Compared with HG group and HG+NC group,the cell activity was significantly in-creased,the amount of LDH released,ROS level and apoptosis rate were decreased,the opening of mPTP was decreased,the expression of caspase-3,caspase-9,Cyt c,Bax,and Apaf-1 was down-regulated,the expression of Bcl-2 was up-regulated,and Bcl-2/Bax ratio was increased in HG+MEG3 group(P＜0.01).Conclusion MEG3 may be involved in the endogenous protective mechanism during hyperglycose-induced neurocyte damage by inhibiting mitochondrion-dependent apoptosis in rats.

Objective To observe the effect of ginkgobalide B (GB) on neurocyte apoptosis and protein kinase B expression in neonatal rats after hypoxic-ischemic brain damage (HIBD).Methods Ninety seven-day-old Sprague-Dawley rats were randomly divided into a sham group,an HIBD group and a GB group,each of 30.HIBD was induced in the HIBD and GB groups using the classical Rice method,while the sham group was given a sham operation.GB (10 mg/kg) was injected intraperitoneally to the rats in the GB group at 0 h and 24 h after the modeling.Then 6 rats were killed 6 h,12 h,24 h and 48 h after the modeling,and the expression of caspase-3 mRNA was detected using a real-time PCR to find the time point of maximum effectiveness.Then to further explore the role of the PI3K-AKT pathway in the anti-apoptosis effect of ginkgolide B,a a GB+LY294002 group of 6 rats,which was injected with PI3K-AKT pathway inhibitor LY294002 (1.8 mg/kg) intraperitoneally at 30 min before the modeling and with GB(10 mg/kg) at 0 h and 24 h after the modeling,was added to the experiment.Hematoxylin-eosin staining,terminal-deoxynucleotidyl transferase-mediated nick end labeling and immunohistochemical staining were then used to observe any morphological changes in the cortex,to detect neuronal apoptosis and to quantify the expression of P-AKT protein.Results The expression of caspase-3 in the HI and GB groups began to increase 6 hours after the HIBD and reached a peak after 24 hours,followed by a gradual decline.The expression of caspase-3 in the GB group was significantly lower than in the HI group throughout,while that of both of those groups was significantly higher than in the sham group.Apoptosis-positive cells and the expression of caspase-3increased had significantly in the HI,GB and GB+LY294002 groups 24 hours after the HIBD compared with the sham group,while the expression of P-AKT protein had decreased significantly.Moreover,the apoptosis-positive cells and the expression of caspase-3 of the HI and GB+LY294002 groups were significantly high-er than those of the GB group,while their expression of P-AKT protein was significantly lower after 24 hours.Conclusion Ginkgobalide B can decrease neurocyte apoptosis caused by hypoxic-ischemic brain damage,especially at 24 h after the damage.The PI3K-AKT signaling pathway plays an important role in this effect.