Objective To explore the effects of different phenotypes macrophages(Mφs)on the proliferation,mi-gration and osteogenic differentiation of periodontal ligament stem cells(PDLSCs).Methods PDLSCs were isola-ted and cultured by tissue block method.Tohoku Hospital Pediatrics-1(THP-1)cell line was stimulated to activate into unpolarized Mφs(M0),then induced to polarize into type Ⅰ Mφs(M1)and type Ⅱ Mφs(M2).Quantitative real-time PCR(qPCR)detected the inflammatory factors tumor necrosis factor α(TNF-α),interleukin(IL)-1 β,IL-6,IL-10 and transforming growth factor-β(TGF-β)mRNA expression level.After collecting culture superna-tants with different phenotypes,PDLSCs were stimulated,native control(NC)group did not receive the culture su-pernatant of Mφs.The effects of PDLSCs proliferation were assessed via Methylthiazolyldiphenyl-tetrazolium bro-mide(MTT)assay,while scratch assays were employed to evaluate their migration.Western blot was utilized to analyze the protein expression of Runt-related transcription factor 2(RUNX2)and alkaline phosphatase(ALP).Additionally,Alizarin Red staining was performed to investigate the deposition of calcified nodules in PDLSCs.Re-sults qPCR showed the relative expression of TNF-α,IL-1 β and IL-6 in M1 Mφs were higher than those in M0 and M2 Mφs(P＜0.05),and the relative expression of IL-10 and TGF-β in M2 Mφs were higher than those in M0 and M1 Mφs(P＜0.05);Western blot showed the expression of RUNX2 and ALP proteins in PDLSCs in M0 and M2 groups was higher than those in the NC group(P＜0.05),Alizarin Red staining showed increased calcified nodule deposition in PDLSCs in M0,M1 and M2 groups compared to the NC group;MTT assay showed the prolifer-ation of PDLSCs in the M0 and M1 groups was suppressed compared to the NC group(P＜0.05);and scratch ex-periment showed the migratory capacity of PDLSCs in the M1 and M2 groups was stronger than that in the NC group.Conclusion M0 and M1 Mφs inhibit PDLSCs proliferation,M1 and M2 Mφs promote PDLSCs migration,and all types of Mφs promote osteogenic differentiation of PDLSCs.

Pseudorabies virus(PRV)is the etiological agent of pseudorabies in pigs,which is char-acterized by dyspnea,reproductive disorders,and neurological diseases,and it spreads widely a-round the world.Since 2011,the newly emerged PRV variants have resulted in poor immunity pro-tection of traditional vaccine strains,and the original method of vaccine strain preparation is time-consuming and labor-intensive.Therefore,it is urgently needed to develop an efficient screening method of the vaccine strain at present.Using CRISPR/Cas9 gene editing technology in this study,two single guide RNAs(sgRNA)were designed targeting the virulence gene TK of PRV variant strain CH/JX/2016,and then the enhanced green fluorescent protein the reporter(EGFP)gene was inserted at the TK locus by a homologous repair plasmid.After multiple rounds of plaque puri-fication,the rPRV-ΔTK/EGFP strain was obtained.The results showed the cleavage efficiency of the two sgRNAs was extremely high.The preparation of rPRV-ΔTK/EGFP strain was succeed af-ter only three rounds of purification,and the EGFP expressed normally.The CRISPR/Cas9 system can edit the PRV gene simply,rapidly,and efficiently,and exhibits great potential in the construction of vaccine candidate strains.Meanwhile,the rescued rPRV-ΔTK/EGFP strain not only could be used as a tracer strain in PRV variant infection progresses,but also for subsequent antivi-ral drug screening.

Objective:To comprehensively evaluated the quality of Sargentodoxae Caulis from different habitats with a combination of indexes and characteristic chromatogram method from Chinese Pharmcopoeia (Edition 2020). Methods:The contents of water content, total ash, ethanolic extract, sulfur dioxide residue, heavy metals and harmful elements, total phenols, chlorogenic acid, salidroside and characteristic chromatogram of 17 batches of Sargentodoxae Caulis were determined. The quality of Sargentodoxae Caulis was comprehensively evaluated by combining chemical pattern recognition method. Results:The water content, total ash content, extracts, and content determination of 17 batches of Sargentodoxae Caulis from different habitats complyed with the provisions of the Chinese Pharmcopoeia (Edition 2020). There were differences in the contents of extracts, chlorogenic acid, and salidroside, among which the content of Anhui origin was higher. A total of 8 common peaks were identified from the 17 batches samples. Conclusion:Comprehensive evaluation of multiple indicators can demonstrate the quality of Sargentodoxae Caulis more correctly, and shows that the quality of Sargentodoxae Caulis from different habitats is different. The quality of Sargentodoxae Caulis from Anhui is better than that from other habitats.

Allergic rhinitis(AR)is a typeⅠallergy of the nasal mucosa mediated by IgE.Allergen specific immunotherapy(AIT)can not only control nasal symptoms effectively and long term,but also change the natural course of disease.Due to the poor re-sponse of some patients to allergen specific immunotherapy and the long course of treatment,it is critical to identify biological indica-tors that can predict and evaluate the efficacy.This review reviews the progress in the study of biomarkers for the prediction and evalua-tion of the efficacy of sublingual immunotherapy(SLIT)for AR.The changes of various immune cells,immunoglobulin and cytokines are related to the efficacy of SLIT and are considered as candidate biomarkers,but their effectiveness and relevance remain to be fur-ther verified.

【Objective】 To explore the mechanism of Yougui Pills in the treatment of knee osteoarthritis （KOA） based on network pharmacology and molecular docking. 【Methods】 The active components of Yougui Pills were obtained by searching TCMSP database. With the active component targets predicted by TCMSP， SwissTargetPrediction and TargetNet， we searched TCMIP， CTD， DisGeNET and GeneCards databases for KOA related targets. By crossing the two sets of targets， we obtained therapeutic targets of Yougui Pills for KOA. FDA-approved drugs for KOA and their targets were searched in the DrugBank database， and the drug-target network was constructed by using Cytoscape. Then the protein-protein interaction （PPI） network was constructed by using STRING database. After Go and KEGG enrichment analyses， the network of “traditional Chinese Medicine-active components-key targets-significant pathways-biological process” was constructed. 【Results】 A total of 132 active components were obtained from the nine herbal medicines of Yougui Pills， corresponding to 490 targets， of which 76 were potential therapeutic targets. Of them 13 were the same as the targets of 5 Western medicines for KOA. We screened out 18 key targets by the PPI network. Go enrichment analysis revealed 1281 biological processes， 60 molecular functions， and 17 cellular components， which were mainly related to inflammation， infection and oxidative stress. KEGG enrichment analysis showed that 50 pathways were significantly enriched， including IL-17 signaling pathway and TNF signaling pathway， whose functions were mainly involved in immune system， signal transduction， endocrine system， cell growth and death and other biological processes. 【Conclusion】 With multiple compounds acting on multiple molecular targets， Yougui Pills can treat KOA through molecular mechanisms including inhibiting inflammation， reducing oxidative stress， and promoting cartilage cell proliferation.

ObjectiveTo investigate the characteristics of polysomnography （PSG） in depression patients complicating moderate-to-severe obstructive sleep apnea hypopnea syndrome （OSAHS）. MethodsA retrospective analysis was conducted on the outpatients， inpatients and physical examination population who completed overnight PSG monitoring in the sleep medicine center of Suzhou Guangji Hospital from December 2017 to October 2019. Four groups of subjects were finally enrolled， including depression patients with moderate-to-severe OSAHS （n=31）， depression patients without OSAHS （n=79）， moderate-to-severe OSAHS patients （n=96） and normal control group （n=32）. The sleep process related indicators （total sleep time， sleep latency， number of awakenings）， sleep structure related indicators （N1， N2， N3， percentage of REM sleep， REM latency， REM sleep duration）， sleep-related respiratory variables （oxygen reduction index） and other polysomnographic parameters of the four groups were compared. ResultsIn terms of sleep process， the total sleep time， sleep latency and number of awakenings yielded significant differences among the four groups （F=2.874， 3.959， 12.291， P<0.05 or 0.01）. In terms of sleep structure， the percentage of total sleep time in N2 and N3 stages demonstrated significant differences among the four groups （F=13.885， 48.013， P<0.01）. The REM latency， REM sleep duration and percentage of REM sleep manifested significant differences among the four groups （F=41.492， 11.827， 10.552， P<0.01）. In terms of sleep-related respiratory variables， the oxygen reduction index exhibited significant differences among the four groups （F=170.585， P<0.05）. ConclusionDepression patients complicating moderate-to-severe OSAHS suffer from severe sleep process and structural disturbances， accompanied by quite frequent and severe sleep-related respiratory events.

Genomic studies of cancer cell alterations,such as mutations,copy number variations(CNVs),and translocations,greatly promote our understanding of the genesis and development of cancers.However,the 3D genome architecture of cancers remains less studied due to the complexity of cancer genomes and technical difficulties.To explore the 3D genome structure in clin-ical lung cancer,we performed Hi-C experiments using paired normal and tumor cells harvested from patients with lung cancer,combining with RNA sequenceing analysis.We demonstrated the feasibility of studying 3D genome of clinical lung cancer samples with a small number of cells(1×104),compared the genome architecture between clinical samples and cell lines of lung cancer,and identified conserved and changed spatial chromatin structures between normal and cancer sam-ples.We also showed that Hi-C data can be used to infer CNVs and point mutations in cancer.By integrating those different types of cancer alterations,we showed significant associations between CNVs,3D genome,and gene expression.We propose that 3D genome mediates the effects of cancer genomic alterations on gene expression through altering regulatory chromatin structures.Our study highlights the importance of analyzing 3D genomes of clinical cancer samples in addition to cancer cell lines and provides an integrative genomic analysis pipeline for future larger-scale studies in lung cancer and other cancers.

Objective:To evaluate the prognostic significance of clonal gene mutations using next-generation sequencing in patients with core-binding factor acute myeloid leukemia (CBF-AML) who achieved first complete remission after induction chemotherapy.Methods:The study, which was conducted from July 2011 to August 2017 in First Affiliated Hospital of Soochow University, comprised 195 newly diagnosed patients with CBF-AML, including 190 patients who achieved first complete remission after induction chemotherapy. The cohort included 134 patients with RUNX1-RUNXIT1 + AML and 56 patients with CBFβ-MYH11 + AML. The cohort age ranged from 15 to 64 years, with a median follow-up of 43.6 months. Overall survival (OS) and disease-free survival (DFS) were assessed by the log-rank test, and the Cox proportional hazards regression model was used to determine the effects of clinical factors and genetic mutations on prognosis. Results:The most common genetic mutations were in KIT (47.6% ) , followed by NRAS (20.0% ) , FLT3 (18.4% ) , ASXL2 (14.3% ) , KRAS (10.7% ) , and ASXL1 (9.7% ) . The most common mutations involved genes affecting tyrosine kinase signaling (76.4% ) , followed by chromatin modifiers (29.7% ) . Among the patients receiving intensive consolidation therapy, the OS tended to be better in patients with CBFβ-MYH11 + AML than in those with RUNX1-RUNXIT1 + AML ( P=0.062) . Gene mutations related to chromatin modification, which were detected only in patients with RUNX1-RUNXIT1 + AML, did not affect DFS ( P=0.557) . The patients with mutations in genes regulating chromatin conformation who received allo-hematopoietic stem cell transplantation (allo-HSCT) achieved the best prognosis. Multivariate analysis identified KIT exon 17 mutations as an independent predictor of inferior DFS in patients with RUNX1-RUNXIT1 + AML ( P<0.001) , and allo-HSCT significantly prolonged DFS in these patients ( P=0.010) . Conclusions:KIT exon 17 mutations might indicate poor prognosis in patients with RUNX1-RUNXIT1 + AML. Allo-HSCT may improve prognosis in these patients, whereas allo-HSCT might also improve prognosis in patients with mutations in genes related to chromatin modifications.

Objective:To explore the clinical effect of Bobath manipulation combined with traditional Chinese medicine fumigation and intradermal acupuncture on shoulder hand syndrome(SHS) after stroke.Methods:From April 2017 to August 2019, 80 patients with SHS after stroke admitted to the People's Hospital of Deqing County were selected, and they were divided into control group and observation group according to the random digital table method, with 40 cases in each group.The control group was treated by Bobath therapy, and the observation group was treated by Bobath therapy combined with acupuncture(intradermal acupuncture) and traditional Chinese medicine fumigation.After 8 weeks of treatment, the ROM scale, FMA scale, BI scale and VAS scores were used to evaluate the improvement in the mobility of the shoulder and wrist joints, motor function of the upper limbs, quality of life and pain, and the clinical efficacy of the two groups was compared.Results:Compared with before treatment, the ROM scale score of all dimensions of shoulder and wrist mobility, FMA scale score and BI scale score of all patients after treatment increased significantly, and the VAS score decreased significantly, the differences were statistically significant(all P<0.05). Compared with the control group after treatment, the improvement of the indicators mentioned above of the observation group were better[shoulder joint flexion ROM score: (154.83±25.63)points vs.(133.82±22.03)points; shoulder joint abduction ROM score: (152.36±25.68)points vs.(133.35±19.96 )points; shoulder joint external rotation ROM score: (75.87±14.69)points vs.(60.82±16.57 )points; wrist joint palm flexion ROM score: (73.94±14.37)points vs.(57.37±9.47)points; wrist joint back extension ROM score: (60.83±7.61)points vs.(42.27±6.37 )points; FMA scale score: (45.74±6.82)points vs.(34.19±4.07)points; BI scale score: (70.36±12.09)points vs.(58.70±12.53)points; VAS score: (1.05±0.49)points vs.(3.37±1.14)points, t=3.703, 3.715, 3.257, 5.576, 7.964, 3.037, 8.746, 3.153, all P<0.05]. The total effective rate of the observation group was significantly higher than that of the control group[97.5%(39/40) vs.62.5%(25/40), χ 2=15.313, P<0.05)]. Conclusion:The combination of Bobath therapy with traditional Chinese medicine fumigation and intradermal acupuncture can improve the pain degree, joint mobility disorder, upper limb motor function and quality of life of SHS patients, the efficacy is better than single Bobath therapy.The clinical effect is accurate, and it is worthy of further promotion and application.

To screen strains with antibacterial and antitumor activity, pregnenolone was used as the sole carbon source for screening bacteria from soil. Based on bacteriostatic activity assay, Pseudomonas aeruginosa HBD-12 was found to be effectively inhibiting the growth of Escherichia coli, Bacillus thuringiensis, Penicillium digitatum and Penicillium italicum, and its fermentation broth was separated and purified using column chromatography. Then, structure of the obtained monomeric compounds was analyzed by spectrum analysis, and their antitumor activity was measured using HTRF kinase detection kit. The isolated monomeric compounds 1-hydroxy-9,10-phenanthroline and 3-hydroxy-9,10-dihydrophenanthroline had significant antitumor activity. At 20 μg/mL, 1-hydroxy-9,10-phenanthroline and 3-hydroxy-9,10-dihydrophenanthroline inhibited 78.39±2.29% and 60.34±8.35% Aurora kinase A, respectively. Therefore, the secondary metabolites of Pseudomonas aeruginosa HBD-12 have the potential to develop antibacterial and antitumor drugs.
Anti-Bacterial Agents/pharmacology* ; Microbial Sensitivity Tests ; Penicillium ; Pseudomonas aeruginosa