Objective To investigate the risk factors for cognitive impairment after ischemic stroke.Metbods The Mini-Mental State Examination (MMSE) scale was used to screen the patients with cognitive impairment within 3 days after the onset of ischemic stroke.The patients were divided into either a cognitive impairment group or a non-cognitive impairment group according to the MMSE scores.Demographics,vascular risk factors,and clinical data were compared in both groups.The independent risks factor for cognitive impairment after ischemic stroke were analyzed by using multivariate logistic regression analysis.Results A total of 202 patients with ischemic stroke were included in the study,in which 48 (23.8％) were in the cognitive impairment group.The proportions of age (66 ± 6 years vs.57 ± 5 years; t =2.231,P =0.038),previous diabetes (39.6％ vs. 18.2％ ; x2 =9.388,P =0.003 ),history of stroke or transient ischemic attack (39.6％ vs.20.8％;x2 =6.856,P=0.007),and the baseline National Institutes of Health Scale scores (11.8 ±2.4 vs,8.1 ± 1.9; t =2.046,P =0.043),as well as serum homocysteine (29.2± 7.8 μmol/L vs.19.9 ±6.5 μmol/L; t =2.781,P =0.008),uric acid (401.5 ± 51.1 μmol/L vs.312.4 ± 60.7 μmol/L; t =3.042,P=0.003),and C-reactive protein (18.4 ±5.2 μmol/L vs.11.3±4.2 μmol/L;t=2.903,P=0.004)levels in the cognitive impairment group were significantly higher than those in the non-cognitive impairment group.Multivariate logistic regression analysis showed that tha age (odds ratio [OR] 1.812,95％ confidence interval [CI] 1.138 -3.205; P =0.039),history of diabetes (OR 2.520,95％ CI 1.854 -4.111; P =0.025),history of stroke or transient ischemic attack (OR 4.232,95％ CI 1.905 - 8.582; P =0.014),as well as the increased levels of serum homocysteine (OR 3.618,95％ CI 2.061 -6.312; P =0.018),uric acid (OR 2.179,95％ CI 1.654 - 3.836; P =0.031),and C-reactive protein (OR 2.716,95％ CI 1.507 - 5.552; P =0.022)were the independent risk factors for cognitive impairment after ischemic stroke.Conclusions The incidence of cognitive impairment after the onset of ischemic stroke was higher.The age and the history of stroke or transient ischemic attack and diabetes,as well as the increased levels of serum C-reactive protein,uric acid,and homocysteine were the independent risk factors for cognitive impairment after ischemic stroke.

Tne voxel-based morphometry(VBM)is a newly developed technique for brain morphometry in recent years.This article briefly expounds the concept,fundamental principle, advantages and disadvantages of VBM.It also reviews the preliminary applications of voxel-based morphometry in mild cognitive impairment and Alzheimer'S disease.

Aim To establish and optimize a method for screening HIV-1 integrase 3′-processing inhibitor. Methods Fluorescence resonance energy transfer ( FRET) was used to create an assay for screening in-tegrase 3′-processing inhibitors; wavelength was de-fined by DNaseⅠ; factors affecting IN activity were optimized, including buffer composition, substrate con-centration, enzyme concentration, metal ion concentra-tion. Results Integrase 3′-processing optimizing reac-tion conditions were buffer 1 , 500 nmol · L-1 sub-strate, 1 μmol·L-1 integrase, 20mmol·L-1 magne-sium ion. Positive drug raltegravir and myricetin could effectively inhibit integrase 3′-processing activity using this assay. Two integrase 3′-processing inhibitors were screened by this method. Conclusion The method for screening HIV-1 integrase 3′-processing inhibitor is successfully established and optimized.

Objective To investigate the relationship between biomarkers of renal function and cerebral microbleeds (CMBs) in cerebral hemorrhage patients.Methods This is a cross-sectional study including a total of 129 patients with cerebral hemorrhage.All patients underwent susceptibility weighted 3.0 T MRI.The presence and number of CMBs on susceptibility weighted MRI were independently interpreted.We calculated the urinary albumin/creatinine ratio (UACR) from morning spot urine and the estimated glomerular filtration rate (eGFR) in serum samples.Serum cystatin C (CysC) was measured using the automated particle-enhanced turbidimetric immunoassay.Results Among 129 patients with cerebral hemorrhage,86 (66.7％) had CMBs on susceptibility-weighted imaging.UACR (mg/g;20.47 ± 9.03 vs 35.24±14.83,t=3.823,P＜0.01)andCysC (mg/L;0.98±0.09vs 1.31 ±0.13,t=4.739,P＜0.01) levels were higher in the patients with CMBs than those without,and the eGFR (ml · min-1 · 1.73 m 2) was lower in the patients with CMBs than those without (78.07 ± 18.69 vs 61.59 ± 17.08,t =3.672,P ＜0.01).A Logistic regression analysis indicated that the levels of kidney impairment biomarkers were significantly associated with the prevalence of CMBs in cerebral hemorrhage patients after an adjustment for age,sex and other risk factors.The odds ratio (OR) and 95％ CI of each kidney biomarkers (eGFR,UACR,and CysC) for the CMBs status were 2.573 (1.172-5.315),2.735 (1.247-6.246)and 2.976 (1.764-6.968),respectively.CysC exhibited fair diagnostic value for CMBs,with an area under the curve of 0.835 (95％ CI 0.791-0.878).Furthermore,there were negative correlations between eGFR and the the number of CMBs (P =0.038,R2 total =0.216).There was a positive correlation between UACR,CysC and number of CMBs (P =0.024,R2 total =0.312;P =0.013,R2 total =0.375).Conclusions Elevated levels of kidney biomarkers are associated with the presence of CMBs in cerebral hemorrhage patients,independent of conventional risk factors.CysC may be a potential diagnostic biomarker for CMBs in cerebral hemorrhage patients.

Objective To study the effect of bone-marrow mesenchymal stem cell (BMSC) via artery transplantation on behavior changes after ischemic brain injury in mice.Methods 60 mice (C57BL/6) were divided randomly into sham group,brain ischemia group (MCAO group) and stem cell therapy group (BMSC group).The latter two groups were injected respectively with 200 μl PBS or BMSC suspension into common carotid arteries namely when removal suture after middle cerebral artery occlusion model,while sham group was only isolated carotid artery.Infarct size of brain tissue was measured by TTC staining.Focal deficit score,Morris water maze test,the rotating beam test and Rotarod test were resepectively made to evaluate the animal behavior after injury.Results Adequate amounts of BMSCs were harvested by adherence screening method and subculture.Ischemic area of BMSC group ((34.98±12.49) ％) was significantly smaller than that of MCAO group ((42.36±9.41)％) at 2nd day after injury (P＜0.05).Compared with MCAO group,focal deficit score of BMSC group reduced significantly at 3rd day after injury,and got to the most significant differences at 5th day after injury (P＜0.01);escape latency of BMSC group was significantly shortened at 7th day and 14th day after injury in Morris water maze test (P＜ 0.05),meanwhile time percentage,distance percentage in the target quadrant and the times corssing the platform were increased gradually after injury,and reached significant differences at 14th day after injury(P＜0.05);exercise time in Rotarod runner increased at every time point after injury,and reached most significant differences at 14th day after injury(P＜0.01);walking speed in the rotating beam test increased most significantly at 14th day after injury,meanwhile walking distance at 5th and 10th day after injury(P＜0.01).Conclusion BMSC transplantation via carotid artery can significantly improve neural function,learning,balance and motor function after brain injury,which will be a new way of TBI therapy.

Objective To comparatively evaluate the effects of a recombinant Mtb protein ESAT 6-CFP10 ( rESAT6-CFP10 ) and a purified protein derivative ( PPD ) as skin test reagents in guinea pigs . Methods Guinea pigs were sensitized with different Mycobacteria species .After sensitization , all guinea pigs were intradermally injected with rESAT6-CFP10 and PPD.At 48 h after the injection, the size of ery-thema at injection sites was measured by using a double-blind method .For guinea pigs sensitized with viable Mtb, the size of erythema at injection sites were measured at 24 h after the injection .The positive conversion rates of skin test with rESAT 6-CFP10 and PPD were calculated .Results The results of PPD skin test were positive in all guinea pigs sensitized with viable Mtb , killed Mtb and BCG with erythema diameters of (11.4 ±0.9) mm, (11.8±1.1) mm and (13.2±0.8) mm, respectively.Positive skin test with rESAT6-CFP10 was only observed in guinea pigs infected by viable Mtb-showing erythema diameters of (13.7±5.7) mm. The skin test with rESAT6-CFP10 was negative in guinea pigs sensitized by killed Mtb-and vaccinated by BCG.The skin tests by using rESAT6-CFP10 and PPD were performed on randomly selected guinea pigs at ninth day after infection by Mycobacterium tuberculosis H37Rv.At the 2nd week, totally 24 selected guinea pigs showed positive skin test results with rESAT6-CFP10 (24/24) with erythema diameters of (19.9± 3.0) mm, while only 15 out of 24 had positive PPD skin test with erythema diameters of (6.1±5.5) mm. At the 4th week, all guinea pigs showed positive PPD skin test (3/3) with erythema diameters of (12.7± 2.5) mm.Conclusion The skin test by using recombinant ESAT 6-CFP10 protein can effectively distin-guish viable Mtb infection from BCG vaccination and killed Mtb sensitization , which is a more suitable anti-gen than PPD for the early diagnosis of Mtb infection .

Objective To investigate the effect of Helicobacter pylori (Hp) infection on plasma lpid and high sensitive C reactive protein (hs-CRP) levels in patients with cerebral infarction, and explore the pathogenesy. Methods Seventy-nine cerebral infarction patients without nearly inflammatory reaction disease were recruited. Hp was detected by breath test,the patients were divided into infection-negative group (15 cases), light infection group (29 cases) and severe infection group(35 cases) according to the results,the last two groups were as infection-positive group. The plasma lipid and hs-CRP levels were exanined.Results Compared with infection-negative group,the total cholesterol,low density lipoprotein,hs-CRP levels increased obviously in infecton-positive group (P ＜ 0.05). The level of hs-CRP in severe infection group was higher than that in light infection group [(10.21 ±4.98) mg/L vs. (5.81 ±4.21) mg/L](P=0.001 ). Conclusions Hp infection may increase vascular inflammatory reaction through lipid metabolic disturbance. The cerebral infarction with higher Hp infection, and with the degree of infection increased, the risk is also increased accordingly.

Objective:To investigate the correlation between heart rate variability (HRV) and cerebral small vessel disease (CSVD) in patients with obstructive sleep apnea (OSA).Methods:Patients with OSA received polysomnography and brain MRI examination in Weihai Municipal Hospital from July 2019 to July 2020 were consecutively collected for cross-sectional analysis. The 5 min HRV before sleep (awake state) was analyzed. The patients were divided into CSVD group and non-CSVD group according to the overall burden of CSVD. The demographic data, clinical data, polysomnography parameters and HRV time domain and frequency domain parameters were compared between the two groups. Multivariate logistic regression analysis was used to determine the correlation between the HRV parameters and CSVD in patients with OSA. Results:A total of 100 patients with OSA were enrolled, including 79 males (79.0%), aged 52.36±8.66 years, apnea hypopnea index (AHI) 38.70±24.65/h. There were 46 patients (46.0%) in the CSVD group and 54 (54.0%) in the non-CSVD group. Univariate analysis showed that there were significant differences in age, AHI, oxygen desaturation index (ODI), percentage of blood oxygen saturation <90% in total sleep time (T90), square root of the mean of the sum of the squares of the difference between adjacent RR intervals (RMSSD), power in high frequency range (HF), power in low frequency range (LF) to HF ratio (LF/HF) between the CSVD group and the non-CSVD group (all P<0.05). Multivariate logistic regression analysis showed that after adjusting for age, body mass index, systolic blood pressure, AHI, ODI, and T90, RMSSD (odds ratio 0.625, 95% confidence interval 0.389-0.981; P=0.041) and LF/HF ratio (odds ratio 1.429, 95% confidence interval 1.011-2.020; P=0.043) were the independent influencing factors of CSVD in patients with OSA. Conclusion:Increased LF/HF and decreased RMSSD in OSA patients with CSVD suggest that the increased sympathetic excitability and decreased vagus function, which may be one of the pathophysiological mechanisms of occurring CSVD in patients with OSA.

Objective:To investigate the correlation between heart rate variability (HRV) and cognitive impairment in patients with obstructive sleep apnea (OSA).Methods:Patients received polysomnography in Weihai Municipal Hospital from June 2019 to November 2020 were enrolled as the subjects of cross-sectional analysis. According to the Montreal Cognitive Assessment score, the patients with OSA were divided into a cognitive impairment group and a non-cognitive impairment group. Multivariate logistic regression analysis was used to determine whether HRV parameters were the independent influencing factors of cognitive impairment in patients with OSA. Multiple linear regression analysis was used to determine the independent correlation between HRV parameters and overall cognition as well as each cognitive domain in patients with OSA. Results:A total of 115 patients with OSA were included, including 80 males (69.6%), aged 58.25±9.88 years. Among them, there were 61 in the cognitive impairment group (53.0%) and 54 in the non-cognitive impairment group (47.0%). The standard deviation of the R-R interval in normal sinus beats (SDNN), the square root of the mean of the sum of the squares of the difference between adjacent NN intervals (RMSSD), the percentage of the number of pairs of adjacent R-R intervals differing by more than 50 ms (pNN50) and the power in high-frequency range (HF; 0.15-0.40 Hz) in the cognitive impairment group were significantly lower than those in non-cognitive impairment group (all P<0.05). Multivariate logistic regression analysis showed that SDNN (odds ratio [ OR] 0.551, 95% confidence interval [ CI] 0.380-0.798; P=0.002), RMSSD ( OR 0.516, 95% CI 0.342-0.779; P=0.002), pNN50 ( OR 0.900, 95% CI 0.834-0.971; P=0.006), LF ( OR 0.821, 95% CI 0.687-0.982; P=0.030) and HF ( OR 0.687, 95% CI 0.525-0.899; P=0.006) were the independent protective factors of cognitive impairment in patients with OSA. Multiple linear regression analysis showed that SDNN ( β=0.208, P=0.023), RMSSD ( β=0.228, P=0.011), pNN50 ( β=0.186, P=0.040), HF ( β=0.235, P=0.010) is independently correlated with overall cognitive function in patients with OSA. Conclusion:The decline of HRV parameters SDNN, RMSSD, pNN50 and HF is independently correlated with cognitive impairment in patients with OSA, suggesting that the decline of vagus nerve function may be involved in the mechanism of cognitive impairment in patients with OSA.

Objective To establish a guinea pig model of latent Mycobacterium tuberculosis infec-tion for evaluating the effects of therapeutic vaccines .Methods Guinea pigs were subcutaneously inocula-ted with 5.0×103 CFU Mtb.The skin test was performed with 0.5μg recombinant ESAT6-CFP10 protein to detect positive conversion rates at different time points .Two weeks after Mtb inoculation , guinea pigs in model group received 5 mg isoniazid treatment ( three times a week for four weeks ) by oral gavage , while those in control group received normal saline .At the sixth week after Mtb infection , guinea pigs with and without isoniazid treatment were dissected for pathology examination .The pathological scores of liver , spleen and lung, as well as bacteria loads in spleen were compared between two groups .The established guinea pig model of latent infection was then validated by testing two reference vaccines ( AEC/BC02 and AEC/BC03 ) . Results Two weeks after Mtb inoculation , all guinea pigs showed positive EC skin test with induration area of (19.9±3.0) mm.Upon four weeks of isoniazid treatment , the guinea pigs in model group showed no pathological changes with zero scores in the examined organs .No bacterium was detected in spleen of ani-mals from model group.However, the total pathological score was 38.8±16.5 and bacteria load in spleen was (5.1±0.3) Log10 CFU with the guinea pigs from control group .Natural recurrence of tuberculosis in model group was observed after drug withdrawal .The total pathological scores were 48.5±23.9 and 51.3± 23.41.The bacterial loads in spleen were (4.5±1.3) and (4.2±1.1) Log10 CFU and bacterial loads in lung were (4.1±1.2) and (3.4±1.3) Log10 CFU respectively as verified with reference vaccines of AEC /BC02 and AEC/BC03.Conclusion Isoniazid treatment inhibited the proliferation of inoculated Mtb in guinea pigs.A guinea pig model of latent Mycobacterium tuberculosis infection is successfully established with an advantage of good repeatability .Therefore, it can be used to evaluate the effects of therapeutic vaccines on latent Mycobacterium tuberculosis infection.