ObjectiveTo investigate the distribution of vascular cognitive impairment （VCI） with kidney Yang deficiency syndrome and explore the biological nature of VCI with kidney Yang deficiency syndrome from the perspective of DNA methylation under the combination of disease and syndrome， so as to provide an epigenetic target for traditional Chinese medicine （TCM） treatment of this disease with this syndrome in the future. MethodCommunity residents in Beijing were screened out for cognitive impairment from September 2020 to November 2022 through the scale， and VCI patients were analyzed for the syndrome. VCI patients with kidney Yang deficiency syndrome and healthy people were enrolled in this study. Peripheral venous blood was collected and subjected to genome-wide DNA methylation detection by Illumina Human Methylation 850K BeadChip. Then， differentially methylated genes （DMGs） were screened out for bioinformatics analysis. ResultA total of 1 902 people were investigated in this study， and 201 of them had VCI， accounting for 10.57%， including 72.14% with kidney Yang deficiency syndrome. The methylation results showed that compared with the normal group， the VCI group had 386 differential methylation sites， and 136 DMGs were annotated. The Kyoto Encyclopedia of Gene and Genomes（KEGG） signaling pathway enrichment analysis showed that the DMGs between the two groups were mainly involved in mammalian target of rapamycin（mTOR） signaling pathway， Estrogen signaling pathway， cyclic adenosine monophosphate（cAMP） signaling pathway， etc. Protein-protein interaction （PPI） analysis showed that DMGs， such as epidermal growth factor receptor（EGFR）， epidermal growth factor （EGF）， and signal transducer and activator of transcription 3（STAT3）， played important roles in the network. ConclusionKidney Yang deficiency is the main syndrome in VCI patients. DMGs including EGFR， EGF， and STAT3 and the related pathways such as mTOR signaling pathway， Estrogen signaling pathway， and cAMP signaling pathway may play a vital role in the occurrence and development of VCI with kidney Yang deficiency syndrome.

Flavanone 3-hydroxylase (F3H) is a key enzyme in the synthesis of phycocyanidins. In this experiment, the petals of red Rhododendron hybridum Hort. at different developmental stages were used as experimental materials. The R. hybridum flavanone 3-hydroxylase (RhF3H) gene was cloned using reverse transcription PCR (RT-PCR) and rapid-amplification of cDNA ends (RACE) techniques, and bioinformatics analyses were performed. Petal RhF3H gene expression at different developmental stages were analyzed by using quantitative real-time polymerase chain reaction (qRT-PCR). A pET-28a-RhF3H prokaryotic expression vector was constructed for the preparation and purification of RhF3H protein. A pCAMBIA1302-RhF3H overexpression vector was constructed for genetic transformation in Arabidopsis thaliana by Agrobacterium-mediated method. The results showed that the R. hybridum Hort. RhF3H gene is 1 245 bp long, with an open reading frame of 1 092 bp, encoding 363 amino acids. It contains a Fe²⁺ binding motif and a 2-ketoglutarate binding motif of the dioxygenase superfamily. Phylogenetic analysis showed that the R. hybridum RhF3H protein is most closely related to the Vaccinium corymbosum F3H protein. qRT-PCR analysis showed that the expression level of the red R. hybridum RhF3H gene tended to increase and then decrease in the petals at different developmental stages, with the highest expression at middle opening stage. The results of the prokaryotic expression showed that the size of the induced protein of the constructed prokaryotic expression vector pET-28a-RhF3H was about 40 kDa, which was similar to the theoretical value. Transgenic RhF3H Arabidopsis thaliana plants were successfully obtained, and PCR identification and β-glucuronidase (GUS) staining demonstrated that the RhF3H gene was integrated into the genome of A. thaliana plants. qRT-PCR, total flavonoid and anthocyanin contentanalysis showed that RhF3H was significantly higher expressed in the transgenic A. thaliana relative to that of the wild type, and its total flavonoid and anthocyanin content were significantly increased. This study provides a theoretical basis for investigating the function of RhF3H gene, as well as for studying the molecular mechanism of flower color in R. simsiib Planch.
Arabidopsis/metabolism* ; Rhododendron/metabolism* ; Amino Acid Sequence ; Anthocyanins/metabolism* ; Phylogeny ; Flavonoids/metabolism* ; Cloning, Molecular ; Gene Expression Regulation, Plant ; Plant Proteins/metabolism*

【Objective】 Compare the early outcome and safety of endoscopy-unilateral laminectomy for bilateral decompression （Endo-ULBD） and posterior lumbar interbody fusion （PLIF） in the treatment of multi-segment lumbar central spinal stenosis. 【Methods】 We retrospectively analyzed 68 patients with multi-segment central lumbar spinal stenosis treated between October 2019 and October 2020 in the Department of Spine Surgery， Affiliated Hospital of Qingdao University. Of them 33 patients were treated with Endo-ULBD and 35 ones were treated with PLIF. We compared the operation time， times of intraoperative fluoroscopy， estimated intraoperative blood loss， incision length， postoperative time to get out of bed， postoperative hospital duration， complications， visual analogue scale （VAS）， Oswestry dysfunction index （ODI） score before and 1 day， 1 month， and 3 months after operation， Japanese Orthopedic Association Assessment Treatment Score （JOA）， and modified MacNab score 3 months after operation between the two groups of patients. 【Results】 Compared with PLIF group， Endo-ULBD group had significantly shorter operation time， smaller incision length， less intraoperative blood loss， shorter postoperative bed time and postoperative hospital stay， and fewer surgical complications （all P<0.05）. There was significantly more intraoperative fluoroscopy in Endo-ULBD group than in PLIF group （P<0.05）. The VAS， ODI and JOA scores of the two groups were significantly improved after treatment （P<0.05）. There was no statistical difference in VAS of leg pain between the two groups after treatment （P>0.05）. However， after treatment Endo-ULBD group outperformed PLIF group in lower back pain VAS， ODI， JOA and the 3-month follow-up excellent and good rates （P<0.05）. 【Conclusion】 For patients with multi-segment central lumbar spinal stenosis， Endo-ULBD treatment can achieve better early clinical outcome than PLIF surgery， with less bleeding， shorter operation time， faster postoperative recovery， and fewer complications.

Objective: To provide guidelines for the prevention of post-stroke dementia, we investigated whether stroke patients were aware of post-stroke dementia and their level of dementia-related knowledge. Methods: Five dementia-related questions were designed. A field survey was conducted in a question-and-answer method using the designed questions. The surveys assessed 3000 stroke patients (2 weeks-6 months after stroke attacks) from 14 hospitals/clinical centers in 7 provinces and cities across China. Results: Among 3000 stroke patients, 80.5％had heard of dementia, 39.1％knew that stroke can lead to dementia, 55.7％ believed dementia can be prevented, 50.8％ thought dementia can be cured, and only 8.8％had ever seen a doctor because of memory deterioration. Then, patients were classified into three groups, including a no cognitive impairment (NCI) group, a mild cognitive impairment (MCI) group, and a mild dementia (MD) group. Among the MCI and MD groups, only 8.7％ (75/861) and 9.9％ (64/649) of patients, respectively, had ever seen a doctor because of memory deterioration. According to our results, patients with a higher level of cognitive impairment had a lower awareness of dementia (P<.001). Conclusion: The awareness of dementia in stroke patients in China is low, and the consultation rate is even lower. Moreover, patients with a higher level of cognitive impairment have a lower awareness of dementia. To improve public awareness and improve prevention, more emphasis should be put on ed-ucation regarding post-stroke dementia. Routine cognitive function screening should be conducted on stroke patients as an effective way to assess dementia.

Objective To explore the association between diabetes mellitus type Ⅱ (DM) and deglutition function of amyotrophic lateral sclerosis (ALS) patients.Methods Seventy-five ALS patients older than 45 years and admitted to our hospital from August 2008 to November 2015 were selected into this study;67 of them were without DM and 8 were with DM.Water swallow test,amyotrophic lateral sclerosis severity scale-swallow (s-ALSSS),amyotrophic lateral sclerosis functional rating scale (ALSFR-R) and videofluoroscopic swallow study (VFS) were performed to evaluate the deglutition functions of these ALS patients with or without DM,and the results of the two groups were compared.Results (1) Patients with DM had significantly higher s-ALSSS scores,ingurgitation part in ALSFRS-R scores and in parts of VFS scores,such as transportation to pharyngeal,pharyngeal transit,flow into pharyngeal before reflexion,epiglottic vallecula residue,and piriform sinus residue than patients without DM (8.88±1.34 vs.7.54±1.47,3.50±0.54 vs.2.96±0.77,2.88±0.35 vs.2.16±0.69,2.75±0.46 vs.2.09±0.69,2.88±0.35 vs.2.42±0.56,2.88±0.35 vs.2.39±0.58,P＜0.05).(2) Scores of Kubota drinking test,ALSFRS-R,VGF (including oral phase,pharyngeal phase and aspiration degree) in patients without DM were 2.15±1.12,7.18±1.41(1.78±0.69,1.69±0.60 and 3.72±0.65),and those in patients with DM were 1.88±1.34,8.00±0.93(2.13±0.64,2.00±0.53 and 4.00±0.00);no significant differences were noted between the two groups (P＞0.05),but there was a trend showing that DM patients had higher scores in these evaluations.Conclusion As compared with ALS patients without DM,ALS patients with DM get more mildly impaired deglutition function.

OBJECTIVE: To purify and identify HMGB1 secreted by liver cells HepG2 and immune cells U937. METHODS: We cultured the liver cell lines HepG2 and immune cell lines U937, and stimulated them with HMGB1 (400 ng/mL) for 20 h. Then the supernatant was collected. Ultrafiltration centrifugation, CM-Sepharose cation, DEAE-Sepharose anion exchange chromatography, Sephadex G75-gel filtration chromatography, and immunoprecipitation were used for purification. The molecular weight and identity of HMGB1 was confirmed by SDS-PAGE and Western blot. RESULTS: A sharp stained protein band with a molecular weight of about 26 kD was obtained by SDS-PAGE analysis and shown to be HMGB1 confirmed by Western blot. CONCLUSION High purified HMGB1 can be separated from these two cell lines.
Cell Culture Techniques ; Electrophoresis, Polyacrylamide Gel ; methods ; HMGB1 Protein ; isolation & purification ; metabolism ; Hep G2 Cells ; Hepatocytes ; metabolism ; Humans ; Monocytes ; metabolism ; U937 Cells

The aim of this study was to investigate the effect of Paris saponin I (PS I) on human gastric carcinoma cell growth and apoptosis and to explore the potential mechanisms. The proliferation of SGC7901 cells was monitored by the MTT cell viability assay, while the nuclear morphology of apoptotic cells was assessed by Hoechst 33258 staining. Flow cytometry was performed to analyze the cell cycle progression of propidium iodide (PI)-stained SGC7901 cells and the apoptotic rate of annexin V/PI-stained cells. Western blotting was used to examine the expression of several cell cycle proteins, including cyclin B1 and Cdk1, and the apoptosis-regulated proteins Bcl-2, Bax, cytochrome c, procaspase-9, and procaspase-3. The MTT assay demonstrated that PS I could induce significant dose- and time-dependent inhibition of SGC7901 cell proliferation. Marked morphological changes, including condensation of chromatin, nuclear fragmentation and apoptotic bodies were clearly shown on Hoechst 33258 staining. PSI treatment also resulted in the disruption of the cell cycle at G(2)/M and the induction of apoptosis. Following PSI treatment, the cell cycle-related proteins cyclin B1 and Cdk1 were down-regulated. Expression of the pro-apoptotic protein Bax was increased, while anti-apoptotic protein Bcl-2 decreased. PSI treatment resulted in elevated cytoplasmic cytochrome c and activation of the apoptotic proteases caspase-9 and caspase-3. These data indicate that PS acts as an inhibitor of proli I feration in SGC7901 cells by inducing cell cycle arrest and mitochondria-dependent apoptosis. PSI is a potential therapeutic agent against human gastric carcinoma.

The aim of this study was to investigate the effect of Paris saponin Ⅰ (PS Ⅰ ) on human gastric carcinoma cell growth and apoptosis and to explore the potential mechanisms.The proliferation of SGC7901 cells was monitored by the MTT cell viability assay,while the nuclear morphology of apoptotic cells was assessed by Hoechst 33258 staining.Flow cytometry was performed to analyze the cell cycle progression of propidium iodide (PI)-stained SGC7901 cells and the apoptotic rate of annexin V/PI-stained cells.Western blotting was used to examine the expression of several cell cycle proteins,including cyclin B1 and Cdkl,and the apoptosis-regulated proteins Bcl-2,Bax,cytochrome c,procaspase-9,and procaspase-3.The MTT assay demonstrated that PSⅠ could induce significant doseand time-dependent inhibition of SGC7901 cell proliferation.Marked morphological changes,including condensation of chromatin,nuclear fragmentation and apoptotic bodies were clearly shown on Hoechst 33258 staining.PSⅠ treatment also resulted in the disruption of the cell cycle at G2/M and the induction of apoptosis.Following PSⅠ treatment,the cell cycle-related proteins cyclin B 1 and Cdk1 were downregulated.Expression of the pro-apoptotic protein Bax was increased,while anti-apoptotic protein Bcl-2decreased.PSⅠ treatment resulted in elevated cytoplasmic cytochrome c and activation of the apoptotic proteases caspase-9 and caspase-3.These data indicate that PSⅠ acts as an inhibitor of proliferation in SGC7901 cells by inducing cell cycle arrest and mitochondria-dependent apoptosis.PSⅠ is a potential therapeutic agent against human gastric carcinoma.

OBJECTIVE To understand the drug-resistant status of Acinetobacter baumanii and provide scientific basis for infection treatment.METHODS Eight multidrug-resistant A.baumannii(MDR-AB) strains isolated from nosocomial inpatients in intensive care unit(ICU) were collected in May 2007 and tested for drug sensitivity and MIC determination as well.We typed A.baumannii isolates with pulsed-field gel electrophoresis(PFGE) to determine whether they derived from the same clone.RESULTS Four isolates from nosocomial inpatients were resistant to multiple antibiotics including carbapenem.The PFGE types identified from eight isolates were same.All A.baumannii isolates are closely related.CONCLUSIONS The prevalence of nosocomial infection is due to transmission of the same strains among the patients in ICU.This finding highly suggests that isolate 1 was the source of this nosocomial infection and the problem of hand hygiene maybe the main transmission route.

OBJECTIVE To investigate the distribution and antimicrobial resistance among clinical isolates of Acinetobacter baumannii(ABA) from July 1 to Dect 31 2007 in Haidian Hospital of Beijing.METHODS A retrospective study on clinical distribution and antimicrobial resistance of 22 antibiotics to ABA was undertaken.RESULTS ABA frequently detected out in phlegm specimen with the detection rate of 86.4%.The drug-resistance to SCF and AMK was the lowest.CONCLUSIONS ABA had severe drug-resistance,SCF and AMK keep better activity to ABA.Rational selection and use of antibacterials is important to prevent the appearance of drug-resistance of ABA.