Objective To investigate the protective effect of nimodipine on neuron of the rats with focal cerebral ischemia-reperfusion injury and the expressions of Bax and Bcl-2,and to clarify their mechanisms.Methods The focal cerebral-ischemia reperfusion model was induced by the middle cerebral artery occlusion(MCAO)method. 30 male Wistar rats were randomly divided into sham operation,model,and nimodipine groups(n=10).The neurological deficit score was performed after 2 h ischemia following 2 h reperfusion.The infarction was observed by TUNEL staining and the expressions of Bax and Bcl-2 were detected by SP immunohistochemistry method. Results Compared with model group, the number of apoptotic cells of the rats in nimodipine group was significantly decreased(P<0.05),the expression of Bax was significantly decreased (P<0.05),and the Bcl-2 expression was increased significantly(P<0.05).The morphological examination showed that the neurons of the rats in model group had serious necrosis and edema while the number of dead cells in nimodipine treatment group was reduced and the edema was improved.Conclusion Nimodipine has a protective effect on brain tissue of the rats with focal cerebral ischemia-reperfusion inj ury, which is closely related to the down-regulation of Bax and up-regulation of Bcl-2 and inhibition of the apoptosis of neuron.

Objective:To analyze the variation and evolution characteristics of hemagglutinin (HA) and neuraminidase (NA) genes of influenza B virus circulating in Jining from 2017 to 2020.Methods:Throat swab specimens were collected from patients with influenza-like symptoms in sentinel hospitals and influenza outbreaks in Jining from 2017 to 2020 and tested for influenza B virus nucleic acid. After virus isolation, 20 representative strains of influenza B virus were selected to sequence the full length of HA and NA genes. Phylogenetic trees were constructed and the molecular characteristics were analyzed using bioinformatics software. Results:A total of 4 575 specimens were collected and 842 of them were positive for influenza virus, including 398 (8.7%, 398/4 575) influenza B virus-positive specimens. The positive rate of influenza B virus was 47.27% (398/842). The isolated influenza B virus strains of Victoria (BV) and Yamagata (BY) lineages from 2017 to 2020 shared 98.7%-98.8% and 98.5%-99.1% homology in HA gene with vaccine strains, respectively. The BV lineage strains isolated from 2018 to 2020 belonged to Victoria clade 1A branch and the BY lineage strains isolated from 2017 to 2018 belonged to Yamagata clade 3 branch. Mutations were detected in several antigenic sites, but not in the sites related to NA inhibitor resistance. Conclusions:Mutations in several antigenic sites caused antigenic changes in influenza B virus of BV and BY lineages, which might be related to the outbreaks of influenza B virus infection in Jining during 2017 to 2020.

Aim To investigate the effect of astragalo-side IV ( ASIV) on myocardial energy metabolism and mitochondrial biosynthesis in myocardial cells of dia-betic rats induced by streptozotocin ( STZ ) . Methods
50 SD rats at 6 weeks of age were assigned to 5 groups,10 for each group:control group, model group, ASIV 10 mg·kg-1 ·d-1 group, ASIV 20 mg·kg-1 ·d-1 group, ASIV 40 mg·kg-1 ·d-1 group. Except the control group,the remaining 40 were used to estab-lish type 1 diabetes model by the tail vein injection of STZ (35 mg·kg-1 ) . At the end of 16 weeks of treat-ment, left ventricular systolic pressure ( LVSP ) , left ventricular diastolic final pressure ( LVEDP ) and left ventricular maximum rising/falling rate ( ± dp/dtmax ) were tested. Pathological section was observed by HE staining. ATP, ADP, AMP levels were detected by ELISA. The expressions of PGC-1α and NRF-1 protein were assessed by Western blot. The expressions of PGC-1α and NRF-1 mRNA were determined by RT-PCR. Results Compared with control group, model group markedly elevated LVEDP and decreased LVSP, ± dp/dtmax , ATP/AMP and ATP/ADP ratio. Com-pared with model group, low-dose ASIV group did not change significantly,middle-dose ASIV group and high-dose ASIV group obviously decreased LVEDP, and im-proved LVSP, ± dp/dtmax , ATP/ADP and ATP/AMP ratio. Meanwhile, the expressions of PGC-1α and NRF-1 protein and mRNA were increased in a dose-de-pendent manner. Conclusion ASIV could promote mitochondrial biosynthesis, improve energy metabolism in myocardial cells of type 1 diabetic rats by PGC-1αand NRF-1 .

Objective To evaluate the effect of breathing exercise program (shrinking lips abdominal breathing combined with vertical breathing gymnastics) on dyspnea, pulmonary function, exercise tolerance and quality of life in patients with moderate to severe chronic obstructive pulmonary disease(COPD). Methods A total of 90 hospitalized patients with moderate to severe COPD were randomly divided into the experimental group and the control group with 45 cases in each group. The experimental group lost one case and the control group lost three cases, 86 patients finished the experiment. During hospitalization, on the base of conventional treatment and care, the experimental group
received training of shrinking lips abdominal breathing combined with vertical breathing gymnastics after medical staff′s care, with folk music as the background. Exercise frequency:morning, afternoon, 2 times per day, 15 min per time and continue to exercise with researchers′ guidance after discharge. In the control group:take exercises by self, take routine follow-up after discharge. The intervention continued for three months. The indicators such as dyspnea, pulmonary function, exercise tolerance and life quality of both groups were assessed respectively before and after the intervention. Results There were no significant differences in dyspnea symptoms, lung function, exercise endurance, quality of life between two groups before intervention (P>0.05). The dyspnea scored 1.43±0.87 after intervention in the experimental group, and 1.93 ±0.97 in the control group, there was significant difference (Z=-2.293, P=0.022). The 6 min walking test distance was (371.34 ± 67.74) m after intervention in the experimental group, and (301.57 ± 61.67) m in the control group, there was significant difference(t = 4.988, P =0.000). The St. George′s Respiratory Questionnaire (respiratory symptoms, limited activity, influence disease) score and total score were 54.73 ± 11.96, 52.55 ± 14.48, 55.45 ± 10.01, 54.56 ± 10.79 after intervention in experimental group respectively, and 61.19 ± 10.72, 61.35 ± 14.66, 60.48 ± 9.39, 60.93 ± 10.16 in the control group, there were significant differences(t=-2.815--2.397, P<0.05). There was no significant difference in lung function after intervention between two groups (P>0.05). Conclusions The program of shrinking lips abdominal breathing combined with vertical breathing gymnastics can relieve dyspnea in patients with moderate to severe COPD, improve exercise tolerance and quality of life, which is a safe and effective rehabilitation for patients with COPD.

Objective:To investigate the mechanism of subjective cognitive decline(SCD)in resting-state by using regional homogeneity(ReHo)and functional connectivity(FC)in SCD patients.Methods:Resting-state functional magnetic resonance imaging(RS-fMRI)was performed in 25 SCD patients and 30 normal controls matched by sex, education and nationality.DPARSFA2.3 and SPM8 software were used to analyze and screen the brain areas with abnormal ReHo values in SCD group, with the posterior cingulated(PCC)/paruneus as seed points for whole-brain FC analysis.Results:Compared with the normal control group, the SCD group showed that ReHo values of right occipital gyrus and left precuneus were increased, and ReHo values of right inferior temporal gyrus, right orbital inferior frontal gyrus and bilateral thalamus were decreased(Voxel level, Alphasim correction, P<0.05). Using PCC/ precuneus as seed voxels, the whole brain functional connectivity analysis showed that the functional connectivity with cerebelum Crus 2 R was increased, and the functional connectivity with right orbital inferior frontal gyrus, left inferior temporal gyrus and temporal pole was reduced(Voxel level, Alphasim correction, all P<0.05). Conclusions:Default mode network may play an important role in the mechanism of SCD, and abnormalities in brain areas may first occur in PCC/precuneus.

Objective To investigate the changes and clinical significance of CD19 ,CD20 ,CD3-CD16+ CD56+ (NK cells) and CD3+CD16+CD56+ (NKT cells) in the children with epilepsy .Methods 125 cases of children with epilepsy children confirmed by Pediatric neurology specialists in Zhongshan People's Hospital from January 2013 to March 2016 were selected as epilepsy group , while 125 cases of healthy children were selected as control group ,and the levels of CD19 ,CD20 ,NK cells and NKT cells in periph-eral blood of two groups were detected by flow cytometry .Results CD19 ,CD20 ,NK cells and NKT cells levels in epilepsy group were higher than those in control group ,and the difference was statistically significant (P<0 .05) .For ROC curve analysis ,CD19 optimal cutoff value of 9 .4 ,the area under the curve was 90 .3% ,with 91 .7% sensitivity .CD20 optimal cutoff value of 7 .75 ,the ar-ea under the curve was 87 .3% ,with 91 .7% sensitivity .NK cells optimal cutoff value of 8 .55 ,the area under the curve was 80 .5% , with 95 .8% sensitivity .NKT cells optimal cutoff value of 0 .35 ,the area under the curve was 76 .9% ,with 91 .7% sensitivity .Con-clusion CD19 ,CD20 ,NK cells and NKT cells can be used as important indexes for monitoring immune conditions in children with epilepsy .

Objective:To explore the correlation between fibular head height and varus knee osteoarthritis occurrence and severity.Methods:A retrospective analysis was performed on 618 participants (618 knees, 184 males and 434 females, mean age 61.12±10.98 years) who underwent standard weight-bearing full-leg radiographs and were diagnosed as non-knee osteoarthritis or varus knee osteoarthritis from January 2019 to June 2019. Knee osteoarthritis was diagnosed according to Kellgren-Lawrence grading: 0-I grades were diagnosed as non-osteoarthritis, II-IV grades were diagnosed as osteoarthritis. Joint line convergence angle (JLCA), medial proximal tibial angle (MPTA) and hip-knee-ankle angle were measured on X-rays to reflect varus deformity. The fibular head height was defined as the vertical distance from upper edge of fibular head to lateral tibial plateau. Patients were divided into 5 groups according to Kellgren-Lawrence grading. Differences of age, gender, height, weight, body mass index, varus deformity (JLCA, MPTA and hip-knee-ankle angle) between Kellgren-Lawrence 0-IV grades were compared. Ordinal logistic regression was performed to analyze the correlation between fibular head height and Kellgren-Lawrence grades. Pearson's correlation analysis was used for the correlation among fibular head height, JLCA, MPTA and hip-knee-ankle angle, and the main factor of JLCA, MPTA and hip-knee-ankle angle was extracted by factor analysis. Multiple linear regressions were used to analyze the correlation between fibular head height and varus deformity score.Results:There were 68, 66, 97, 98, 289 participants in Kellgren-Lawrence grades 0-IV respectively that was 134 participants were diagnosed as non-osteoarthritis and 484 participants were diagnosed as osteoarthritis. Fibular head height and MPTA showed a decreasing trend ( F=129.076, 24.875; P<0.001) while JLCA and hip-knee-ankle angle showed an increasing trend ( F=414.346, 105.996; P<0.001) with the increase in Kellgren-Lawrence grading. Age, body mass index and fibular head height are influencing factors of Kellgren-Lawrence grading with OR(95%CI) were 1.116(1.093, 1.141), 1.363(1.060, 1.754), 0.617(0.575, 0.662) . Fibular head height was negatively correlated with JLCA and hip-knee-ankle angle ( r=-0.641, -0.478; P<0.001) , respectively, and positively correlated with MPTA ( r=0.320, P<0.001). There were significant correlations between age, fibular head height and the varus deformity score ( β=0.274, -0.457; P<0.001). Conclusion:Fibular head height of patients with varus knee osteoarthritis is lower than that of non-osteoarthritis. In addition to age and body mass index, fibular head height is a risk factor for varus knee osteoarthritis occurrence. The smaller the fibular head height is, the more serious the osteoarthritis severity and varus deformity are.

The objective of the study was to clone avian beta-defensin (AvBD) 5 gene from pigeon bone marrow tissues and liver tissues, to express the recombinant AvBD5 protein in E. coli, and to determine its antimicrobial activity. The mRNA of duck AvBD5 was cloned from pigeon bone marrow tissues and liver tissues by RT-PCR. In addition, phylogenetic relationships between amino acid sequence of the pigeon AvBD5, AvBDs from other avian species, and some mammalian beta-defensin-5 were analyzed. The cDNA of pigeon AvBD5 was sub-cloned into pGEX-6p-1 vector to construct recombinant plasmid pGEX-pigeon AvBD5. The recombinant protein was expressed into E. coli and purified. Antimicrobial activity and physical-chemical stability of the recombinant fusion protein were measured in vitro. The complete nucleotide sequence of both cDNAs contained 201 bp nucleotides, encoding a polypeptide of 66 amino acids. Both beta-defensins have six conserved cysteines. Phylogenetic relationships were analyzed. Both pigeon AvBDs shared the highest amino acid homology (87.9% and 78.8%) with duck AvBD5. So it was named as pigeon AvBD5alpha (bone marrow) and AvBD5beta (liver). Both recombinant plasmids were transformed into E. coli BL21 and the bacteria were induced with Isopropyl beta-D-1-Thiogalactopyranoside (IPTG). After purification, antibacterial activity of the purified was investigated. In addition, effect of ionic strength on the antibacterial activity, and hemolytic recombinant protein activity of the purified recombinant protein were investigated. A 32 kDa protein was highly expressed. Both purified recombinant pigeon AvBD5alpha and AvBD5beta exhibited extensive antimicrobial activities against 12 bacteria, including Gram-positive and Gram-negative. In high salt ions concentrations, antibacterial activity of both recombinant proteins was decreased. In addition, the hemolysis activity of recombinant protein was extremely low.
Amino Acid Sequence ; Animals ; Anti-Infective Agents ; metabolism ; pharmacology ; Avian Proteins ; biosynthesis ; genetics ; pharmacology ; Cloning, Molecular ; Columbidae ; genetics ; Escherichia coli ; genetics ; metabolism ; Molecular Sequence Data ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; beta-Defensins ; biosynthesis ; genetics ; pharmacology

Objective:To investigate regional homogeneity(ReHo)and whole brain functional connectivity(FC)in patients with mild cognitive impairment(MCI), and to explore the mechanisms of MCI in the resting state.Methods:Resting-state functional magnetic resonance imaging(RS-fMRI)was performed on 24 patients with MCI and 30 age, gender and nationality-matched normal controls.Abnormal brain areas in the MCI group were screened and analyzed by using the DPARSFA2.3 and SPM8 software programs.Whole brain FC analysis was performed with the posterior cingulate cortex(PCC)/precuneus as the seed points.Results:Compared with the control group, MCI subjects displayed higher ReHo values in the frontal-middle-Left, precentral-Left, postcentral-Left, rolandic-opercular-Left, and frontal-inferior-opercular-Left and lower ReHo values in the temporal-superior-right, temporal-middle-right, postcentral-right, and temporal -pole -superior -right(Voxel level, Alphasim correction, P<0.05). Whole brain FC analysis showed greater functional connectivity of PCC/precuneus with fusiform-right, thalamus-right, lingual-right and parahippocampal-right in subjects with MCI, and less functional connectivity of the PCC/precuneus with temporal-middle-Left, angular-Left, temporal-superior-Left and occipital-middle-Left in subjects with MCI(Voxel level, Alphasim correction, P<0.05). Conclusions:Abnormalities of the default mode network may be associated with the onset of MCI, and abnormalities in posterior cingulate/precuneus connectivity may be helpful in finding imaging evidence with high sensitivity to MCI.

Objective:To investigate the characteristics of the nocturnal melatonin secretion concentration and circadian rhythm in patients with wake-up stroke (WUS).Methods:Patients with acute ischemia stroke (AIS) admitted to the Department of Neurology, the Second Affiliated Hospital of Soochow University from October 2019 to August 2022 were enrolled. They were divided into WUS group and non-WUS group. Saliva samples within one week after admission were collected (at 19∶00, 20∶00, 21∶00, 22∶00, and 23∶00) and melatonin concentration was measured. Melatonin secretion curve graph was drawn, dim light melatonin onset (DLMO) was calculated, and circadian rhythms were evaluated. The differences in endogenous circadian rhythms between the WUS group and the non-WUS group were compared. The relevant factors of WUS were determined by multivariate logistic regression analysis. Results:A total of 116 patients with AIS were included, with 79 males (68.1%), aged 59.9±10.3 years; 35 patients (30.2%) were WUS. Univariate analysis showed that there was a statistically significant difference in the infarct site between the WUS group and the non-WUS group ( P=0.019). At 21:00 ( P=0.004) and the average ( P=0.038) nighttime melatonin concentration in the WUS group were significantly lower than those in the non-WUS group, and DLMO showed a significant delay compared to the non-WUS group (21:28:08 vs. 20:57:57; P=0.015). Multivariate logistic regression analysis showed a significant independent correlation between DLMO delay and WUS (odds ratio 1.792, 95% confidence interval 1.123-2.858; P=0.014). Conclusion:Patients with WUS may have endogenous circadian rhythm delay, which is an independent risk factor for WUS.