Objective To investigate the dynamic changes of ATP content in rat cerebral cortex after transient ischemia followed by reperfusion and the relationship between the change of energy and the recovery of neural function.Methods The rats were subjected to 10 min of middle cerebral artery occlusion (MCAO). At the time point of 0 h, 1 h, 3 h, 6 h, 12 h, 24 h and 72 h after reperfusion, ATP contents of frontal and parietal cortex were measured by capillary zone electrophoresis.Results At the end of 10 min ischemia, ATP content fell dramatically to less than 20% of the control level. After reperfusion, ATP content recovered gradually. After 1 h, 3 h, 6 h and 12 h of reperfusion, ATP content returned to 70.5%, 65.7%, 84.8% and 86.9% of the control level ( P=0.052, 0.030, 0.332 and 0.491). From 24 h on until 72 h after reperfusion, ATP content decreased again, reaching half of the control level ( P=0.003 and P=0.023). After 10 min ischemia, limb function recovered gradually and completely at last. From 24 h on until 72 h after reperfusion, unwillingness of action and eating was found.Conclusions The recovery of cellular energy system function is delayed even though the reperfusion is in time after transient cerebral ischemia. Furthermore, secondary failure of cellular energy system function occurrs with the reperfusion proceeding. These phenomena are probably responsible for the delayed recovery of neural function after cerebral ischemia in spite of reperfusion.

Objective To explore the effect of 14,15-epoxyeicosatrienoic acids (14,15-EET) on the inflammatory response of BV2 cells under oxygen and glucose depriviation/reoxygenation (OGD/R) conditions.Methods BV2 cells were randomly divided into three groups,blank control group,vehicle control group,and 14,15-EET group.Under treatment of 14,15-EET,the concentration of inflammatory factor in BV2 cell culture media was detected by ELISA at different time points (reoxygenation for 0,3,6,12,24 h) after OGD1h.The viability of BV2 cells was detected by MTT assay at different time points.At the same conditions,using Transwell migration experiment,migration ability of BV2 cells was observed.Results The 14,15-EET group had the lower levels of inflammatory factor secretion,lower viability and weaker ability of migration than the vehicle control group.The above results were most statistically significant at OGD1h/R12h.Conclusion 14,15-EET can inhibit the inflammation of BV2 cells induced by the injury of OGD reperfusion.

This study was aimed to examine the effect of TREK-1 silencing on the function of astrocytes. Three 21-nucleotide small interfering RNA (siRNA) duplexes (siT1, siT2, siT3) targeting TREK-1 were constructed. Cy3-labeled dsRNA oligmers were used to determine the transfection efficiency in cultured astrocytes. TREK-1-specific siRNA duplexes (siT1, siT2, siT3) at the optimal concentration were transfected into cultured astrocytes, and the most efficient siRNA was identified by the method of immunocytochemical staining and Western blotting. The proliferation of astrocytes tranfected with TREK-1-targeting siRNA under hypoxia condition was measured by fluorescence-activated cell sorting (FACS). The results showed that TREK-1 was expressed in cultured astrocytes. The dsRNA oligmers targeting TREK-1 could be transfected efficiently in cultured astrocytes and down-regulate the expression of TREK-1 in astrocytes. Moreover, the down-regulation of TREK-1 in astrocytes contributed to the proliferation of astrocytes under hypoxia condition as determined by cell cycle analysis. It was concluded that siRNA is a powerful technique that can be used to knockdown the expression of TREK-1 in astrocytes, which helps further investigate the function of TREK-1 channel in astrocytes under physicological and pathological condition.

Objective To investigate the association of the polymorphism of rs3750344 and rs1435252 of G-protein family GABABR2 gene with obesity in population of Uygur and Han. Methods 785 Uygur subjects from Xinjiang and 425 Han subjects from Jinan Maternity and Child Care Hospital were recruited by epidemiology survey. Exposure indicators such as body mass index (BMI), total cholesterol (Tch), triglyceride (TG), fasting glucose (Glu) concentration were measured. Two single nucleotide polymorphisms (SNPs) of rs3750344 and rs1435252 of GABABR2 gene were typed with Taqman. Linkage disequilibrium and haplotype were analysed with Haploview software. Results The frequency of rs1435252 was significantly different in AA carries of Uygur subjects between overweight group (12.5%) and normal weight group (6.6%), the subject with AA genotype significantly increased risk of overweight (OR=1.43, 95% CI: 1.06~1.91). The frequency of rs1435252 was significantly different in A alleles carries of Han subjects between obesity group (80.0%) and normal weight group (65.2%), the subject with A alleles significantly increased risk of obesity (OR=2.13, 95%CI: 1.18~3.86). The C alleles of rs3750344 significantly decreased risk of obesity (OR=0.69, 95%CI: 0.49~0.97) in Uygur, but the significance disappeared after controlling for covariates of age and gender. Conclusion The rs1435252 A allele of GABABR2 gene is a risk factor for overweight or obesity in population of Uygur and Han.

Objective To evaluate the imaging diagnosis of anomalous origin of coronary artery from the pulmonary artery(ACAPA).Methods A total of 11 cases with ACAPA were included in the present study.Chest films,echocardiography,cardioangiography,and electron beam computed tomography (EBCT) were employed as diagnostic modalities.Macroscopic anatomy at operation was referred. Results Ten cases were classified as anomalous origin of left coronary artery from the pulmonary artery(ALCAPA) and 1 case as anomalous origin of right coronary artery from the pulmonary artery(ARCAPA).They could not be diagnosed by chest films,but could be diagnosed by echocardiography in 3 cases,by EBCT in 1 case,and by cardioangiography in all cases.In ALCAPA,cardioangiography showed that the left coronary arteries arising from the posterior sinus or posterior wall of the pulmonary artery were perfused retrogradely via the collaterals from the dilated right coronary artery.In ARCAPA,the right coronary artery originated from the right sinus of the pulmonary artery.Gross anatomy at operation showed that the sites of the anomalous origins were the same as that of cardioangiography.Ischemic fibrosis of the anterior papillary muscles,mitral valve annulus enlargement,and prolapse of mitral valve,which led to mitral valve insufficiency,were found in 3 cases.Conclusion Chest film has limitation in the diagnosis and echocardiography should be further improved.Cardioangiography remains the “gold standard” of the preoperative diagnosis.

AIM: To investigate effects of different therapeutic methods and typical recipes on activation of ERK1/2 in Kupffer cells of rats with fatty liver.METHODS: The rat model of fatty liver was established by feeding high fat diet combinated with distillate spirit.Meanwhile Chinese medicines Shugan fang,Jianpi fang,Huoxue fang,Qushi fang,and Zonghe fang were given to treat different groups respectively.12 weeks later,the Kupffer cells were isolated from livers of control group,model group and different treatment groups by sequential in situ perfusion with collagenaseⅣ and pronase E,density gradient centrifugation,selective adherence.The expression of total ERK1/2 and phospho-ERK1/2 in Kupffer cells of control group,model group and different treatment groups were detected by Western blotting.RESULTS: The expressions of total ERK1/2 and phospho-ERK1/2 were higher in Kupffer cells from model group than those in control group(P

This study was aimed to examine the effect of TREK-1 silencing on the function of astrocytes. Three 21-nucleotide small interfering RNA (siRNA) duplexes (siT1, siT2, siT3) targeting TREK-1 were constructed. Cy3-labeled dsRNA oligmers were used to determine the transfection efficiency in cultured astrocytes. TREK-1-specific siRNA duplexes (siT1, siT2, siT3) at the optimal concentration were transfected into cultured astrocytes, and the most efficient siRNA was identified by the method of immunocytochemical staining and Western blotting. The proliferation of astrocytes tranfected with TREK-1-targeting siRNA under hypoxia condition was measured by fluorescence-activated cell sorting (FACS). The results showed that TREK-1 was expressed in cultured astrocytes. The dsRNA oligmers targeting TREK-1 could be transfected efficiently in cultured astrocytes and down-regulate the expression of TREK-1 in astrocytes. Moreover, the down-regulation of TREK-1 in astrocytes contributed to the proliferation of astrocytes under hypoxia condition as determined by cell cycle analysis. It was concluded that siRNA is a powerful technique that can be used to knockdown the expression of TREK-1 in astrocytes, which helps further investigate the function of TREK-1 channel in astrocytes under physicological and pathological condition.
Animals ; Astrocytes ; physiology ; Cells, Cultured ; Gene Silencing ; physiology ; Potassium Channels ; Potassium Channels, Tandem Pore Domain ; genetics ; RNA Interference ; physiology ; RNA, Small Interfering ; genetics ; Rats

Objective: To investigate the roles of N-acetyl-L-cysteine (NAC) against binge drinking-induced fatty liver in mice. Methods: SPF male C57BL/6 mice were randomly divided into 3 groups, i.e. control group, model group, and NAC/ethanol group (n=10). Mice in model and NAC/ethanol groups were exposed to 3 doses of ethanol (6 g/kg bw) to induced fatty liver, while mice in control group received equal volume and equal energy of maltodextrin solution. NAC was administered to mice at 1 h before ethanol exposure (100 mg/kg bw, i.p.). The mice were sacrificed at 6 h after the last ethanol exposure. The liver and epididymal adipose tissues were collected. Histopathological examination and biochemical assay kit were used to evaluate the fat accumulation, while Western-blot was performed to detect the protein levels of some key factors involved in fat metabolism in liver and adipose tissues. Results: Compored with control group mice, the liver index and liver weight were significantly increased compared with model group, the liver index and TG level in NAC/ethanol group mice were all significantly decreased (P<0.05). Histological examination showed NAC effectively suppressed binge drinking-induced fat accumulation in mice liver. In addition, NAC had no significant effects on the protein levels of peroxisome proliferator-activated receptor-α (PPAR-α), Acy-CoA oxidase (ACOX), sterol regulatory element binding protein 1 c (SREBP-1c) and fatty acid synthase (FAS). Furthermore, the protein levels of hormone sensitive lipase (HSL) did not significantly differ among 3 groups, whereas NAC prevented binge drinking-induced increase of HSL phosphorylation at ser563 and ser660. Conclusion NAC could effectively attenuate binge drinking-induced fatty liver, which might be associated with the inhibition of lipid mobilization by suppressing the phosphorylation of HSL.

Objective:To assess the validity and reliability of the depression and somatic symptoms scale among patients with coronary heart disease.Methods:Totally 246 patients with coronary heart disease were assessed with depression and somatic symptoms scale (DSSS), Hamilton depression rating scale for depression (HAMD) and patients’ health questionnaire depression scale-9 item (PHQ-9). The structural validity was evaluated with exploratory factor analysis and confirmatory factor analysis. The validity as a screening tool was evaluated with the gold standard diagnosed by psychiatrists who were trained with the mini international neuropsychological interview (MINI) according to ICD-10. Receiver operating characteristic (ROC) curve was used to identify cutoff scores for depression. Cronbach α coefficient was used to evaluate the internal consistency.Results:Exploratory factor analysis yielded two factors: depression factor and somatic factor, and the cumulative variance was 51.8%. The fitting indexes of confirmatory factor analysis were as follows: χ2/ df=3.636, RMR=0.077, RMSEA=0.104, IFI=0.804, TLI=0.781, CFI=0.802. The intraclass correlation coefficient of DSSS and HAMD was 0.54. The area under ROC curve (AUC) was 0.828, and the best boundary value was 17 points (sensitivity and specificity: 81% and 75%, respectively). The total scores and subscale scores for internal consistency of DSSS were higher in the depression group than those in the non-depression group ( P<0.01). Cronbach α coefficient for internal consistency of DSSS was 0.917. Conclusion:The DSSS has good validity and reliability among patients with coronary heart disease for screening depression, and can be used to screen depression among patients with coronary heart disease in general hospital.

Objective:To investigate the nutritional risk and prevalence of malnutrition in patients with terminal stage gastrointestinal malignant tumors in a tertiary hospital in Changsha.Methods:Cluster sampling was used to conduct a cross-sectional survey of inpatients from Departments of Gastroenterology, Gastrointestinal Surgery, Hepatobiliary Surgery and Oncology in Hunan Provincial People's Hospital from January 2019 to July 2020. Nutritional Risk Screening 2002 (NRS 2002) was used to assess the prevalence of nutritional risk with malnutrition defined as concurrent presence of BMI < 18.5 kg/m 2, poor general condition and NRS 2002 nutritional impairment score of 3. Step 2 of Global Leadership Initiative on Malnutrition (GLIM) diagnostic criteria (without whole body muscle mass) was adopted to diagnose malnutrition. Step 3 of GLIM criteria was used to evaluate the prevalence of severe malnutrition. Results:A total of 802 patients registered in the 4 departments were selected for screening via cluster sampling and 514 were enrolled according to the inclusion/exclusion criteria. The prevalence of nutritional risk in patients with terminal stage gastrointestinal cancer was 49.8% (256/514). The prevalence of malnutrition and severe malnutrition per GLIM criteria were 41.6% (214/514) and 18.3% (94/514), respectively.Conclusions:Although nutritional support therapy is not recommended for patients with end-stage cancer. This paper suggests that the prevalence of nutritional risk and malnutrition in patients with end-stage gastrointestinal cancer is not as high as described in some articles.