A breakthrough in molecular biology for the twenty-first century is CRISPR/Cas gene editing, which has been used in a variety of fields due to its simplicity, adaptability, and targeting. Given the current global challenge of severe bacterial resistance, difficulties in detecting antimicrobial resistance, and slow development of antimicrobial drugs, CRISPR/Cas gene-editing technology offers a promising avenue for the development of antibacterial treatments. On the one hand, CRISPR/Cas gene editing technology helps advance the study of bacterial functions and serves as a toolbox. For instance, Cas proteins and exogenous repair systems enable efficient and precise gene editing, nCas proteins and deaminase systems facilitate template-free and single base precision editing, dCas proteins and reverse transcriptase allow for repair-free gene editing, and dCas proteins and modified sgRNA enable gene expression level regulation and gene function analysis. On the other hand, its specific gene recognition and targeted DNA cleavage characteristics can be used for pathogen detection, elimination of drug-resistant bacteria and genes, and hold promise as a new strategy for clinical diagnosis and treatment.

Adult ; Aged ; Antihypertensive Agents ; therapeutic use ; Capsules ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Hypertension ; drug therapy ; Male ; Middle Aged ; Phytotherapy

Bone Marrow Cells ; metabolism ; DNA-Activated Protein Kinase ; genetics ; metabolism ; Humans ; Leukemia ; genetics ; Nuclear Proteins ; genetics ; metabolism ; PTEN Phosphohydrolase ; genetics ; metabolism ; RNA, Messenger ; Telomerase ; genetics ; metabolism

Objective To study the expression and significance of monocyte chemotactic protein-1 (MCP-1/CCL2) and vascular endothelial growth factor (VEGF) in serum samples of oral squamous cell carcinoma (OSCC) patients.Methods The concentrations of CCL2 and VEGF in the serum was assessed by ELISA in healthy donors (n=27) and OSCC patients (n=85).Then analyzed the correlation between the concentrations of CCL2 and VEGF and the relationship with patients' clinicopathological characteristics.Results CCL2 concentration was lower in OSCC patients than in healthy donors (69.12 ± 19.54 pg/ml vs 103.41 ± 34.42 pg/ml,t =6.477,P＜0.05).The expression of CCL2 was positively associated to TNM stage in OSCC (t=2.193,P＜0.05).VEGF concentration was higher in OSCC patients than in healthy donors (145.76 ± 49.34 pg/ml vs 70.35± 14.93 pg/ml,t=3.92,P＜0.05).There was a negative correlation between CCL2 and VEGF (r=-0.216,P＜0.05).The receiver operating characteristic (ROC) curve suggests that CCL2 and CCL2/VEGF in serum are good diagnostic markers to discriminate healthy people from OSCC patients,the cutoff values was 98.61 pg/ml and 0.82.Conclusion The expression of CCL2 and VEGF in serum correlated to OSCC progression,and it can be a potential diagnostic biomarker for oral disease.

With comparison of three different methods for the marking of amines compound, an optimal deri vatization method was selected.5-(2-Hydroxyethyl) benzoacridine (HBA) reacts with coupling agent N,N'-carbonyldiimidazole(CDI) to form an activated amide intermediate 2-(benzoacridin) ethyl-imidazole-1-carbox-ylate(BAEIC).BAEIC, which is dual-sensitive probe, reacts preferably with amino compounds at 80 ℃ in the presence of 4-dimethylaminopyridine(DMAP) catalyst in N,N-dimethylformamide(DMF) solvent to give the corresponding sensitively fluorescent derivatives with an excitation maximum at λ_(ex) of 280 nm and an emis sion maximum at λ_(em) of 510 nm.BAEIC-amine derivatives simultaneously exhibited high ionization potential with percent ionization (changing from 5.62% to 58.08% in aqueous acetonitrile and from 2.14% to 56.58% in aqueous methanol.Derivatives were not only sensitive to fluorescence but also to MS ionizable potential.The fluorescence detection limits(5/iV = 3) were 0.12-0.59 μg/L.The online APCI-MS detection limits were 1.9-14 μg/L(S/N=5).

Objective:To identify the risk factors of anxiety disorders among patients undergoing coronary angiography (CAG) and to determine whether the decision of revascularization affect anxiety level following coronary angiography.
Methods:A total of 379 patients undergoing CAG in Fuwai Hospital from Dec. 2012 to Dec. 2013 were invited to participate this study. A data-collecting form, which included questions about demographic features, health history, Type A Behavior Questionnaire (TABQ) and Self Rating Anxiety Scale (SAS), was completed by the participants on the day before and the day after CAG.
Result:Among these patients, SAS score of both before and the day after CAG were higher than Chinese normative SAS score. Female patients had a higher SAS score level than male patients (40.57±9.53 vs 38.26±9.61, P<0.05) before CAG. Patients with these factors of female, over 50 years old, duration of coronary artery disease over 1 year, lower education level had a higher SAS score after CAG. SAS score declined signiifcantly after CAG except those scheduled to receive CABG. Multivariable linear analysis found the D-value between SAS scores before and after CAG was negative correlated with degree of education and positive correlated with the duration of coronary artery disease.
Conclusion:The anxiety level decreased after CAG, except those who need coronary revascularization surgery.

Objective An effective reproducible protocol for complete plant regeneration via somatic embryogenesis has beendeveloped for Herpetospermum pedunculosum,an endangered Tibetan medicinal herb.Methods The cotyledonexplants used in this study were excised from seedlings germinated in vitro.Callus was induced from cotyledonexplants on Murashige and Skoog's medium,supplemented with 2,4-dichlorophenoxyacetic acid(2,4-D,0.1-1.0mg/L)alone or in combination with 6-benzylaminopurine(BA,0.5,1.0,and 2.0 mg/L).Results The calli showeddifferentiation of globular embryos after three weeks of incubation on MS medium supplemented with variouscombinations of BA and NAA.Sixty-two percent of the embryogenic calli produced somatic embryos in MS basalmedium supplemented with BA(1.0 mg/L)+NAA(2.0 mg/L).The addition of KN(0.5 mg/L)to MS mediumcontaining both BA and NAA(2.0 mg/L each)significantly increased the frequency of somatic embryogenesis.Themaximum percentage of embryogenic calli formation was 83%,and globular embryos formed and germinatedsuccessfully in this medium.Then,transferring the regenerated plants from this medium to hormone-free MSmedium will further enhanced the development of the plants,and the healthy plantlets are formed successfullywithin four weeks.The plantlets were transferred to soil to acclimatize under greenhouse conditions and 75%survived.Conclusion Somatic embryogenesis protocol as reported here can play a key role in the propagation andconservation of this endangered species.

AIMTo study the chemical composition in the fruits of Liquidambar formana Hance.

METHODSVarious column chromatographic techniques were used to separate and purify the constituents. Their physical and chemical properties and spectral data were measured for structural elucidation.

RESULTSEleven compounds were isolated and identified as beta-sitosterol (1), 3-oxo-11 alpha, 12 alpha-epoxyleanan-28, 13 beta-olide (2), 3-oxo-12 alpha-hydroxy-oleanan-28, 13 beta-olide (3), 3 alpha-acetyloxy-25-hydroxyolean-12-en-28-oic acid (4), oleanolic acid (5), ursolic acid (6), daucosterol (7), betulonic acid (8), gallic acid (9), nonacosane (10) and n-triacontanoic acid (11).

CONCLUSIONAmong the isolated constituents, compound 4 is new compound, compound 3 is firstly isolated from the natural product and compound 5, 6, 7, 9, 10 and 11 are isolated from LuLuTong for the first time.

Fruit ; chemistry ; Liquidambar ; chemistry ; Molecular Conformation ; Molecular Structure ; Oleanolic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry

ObjectiveToinvestigatetheinflammatoryresponseandapoptosisof peripheral blood mononuclear cells(PBMCs) and their regulation by triptolide(TP) in IgA nephropathy(IgAN) patients.MethodsBlood samples were collected from 29 IgAN patients and 16 healthy individuals.TNF-α and IL-6 concentrations were measured by ELISA and NO concentration by Griess reagent in the plasma of samples.PBMCs were isolated from IgAN patients and cultured in vitro,and subsequently activated by PHA(10 mg/L).The cytotoxicity of different TP concentrations was assayed by MTT and two non-toxic concentrations (12.5 μg/L or 25.0 μg/L)were selected for treatment.TNF-α,IL-6 and NO concentrations were measured in the culture media collected from PBMCs cultures activated by PHA (10 mg/L) and treated with TP (12.5 μg/L or 25.0 μg/L).The PHA-activated,TP-treated cells apoptotic rate was analyzed by FACS using Annexin V-FITC staining.The expression of Bcl-2,Bax,caspase-9 and caspase-3 were detected by RT-PCR and Western blotting from lyses of PHA-activated with or without TP-treated cells.ResultsThe serum concentrations of TNF-α[(131.57±50.61) ng/L vs(30.24±18.93) ng/L,P＜0.01],IL-6[(76.36±25.21) ng/L vs(35.08±16.59) ng/L,P＜0.01] and NO[(46.36±12.93) μmol/Lvs (26.61 ±10.87) μmol/L,P＜0.01] were significantly increased in IgAN patients compared to healthy individuals.PBMCs viability in culture decreased after TP treatment in a dose-dependent manner.TP also inhibited TNF-α,IL-6 and NO levels in the media of PHA-activated PBMCs in culture and induced PBMCs apoptosis.The expression of Bcl-2 decreased markedly and Bax,caspase-9 and caspase-3 increased significantly after TP treatment (all P＜0.05).Conclusions The PBMCs from IgAN patients are in a highly activated state,and have a high apoptotic rate.TP treatment induces benificial effects in IgAN patients by inhibiting the activation of PBMCs by activating pro-apoptotic pathway.

To understand Lophatherum gracile plant community's structural characteristics, a survey of community structure and species diversity was conducted through quadrat sampling in Yongchuan district of Chongqing. The results showed that there were 386 species vascular plants, belonging to 117 families and 229 genera. Based on habitat, community structure and species composition, L. gracile were found in three community types: Pinus massoniana community, banboo community, shurb community. Vertical structure was composed of three layers, including tree layer, shrub layer and herb layer. Species in shrub layer was the richness. P. massoniana is the only dominant species of the community, it can not regenerate naturally, the shrub layer has a greater effect on the community of L. gracile in the future. In addition, the banboo community and shurb community is not stable because of human's activity. Therefore, the community characters of L. gracile should be taken care of conservation when the resources are utilized.
China ; Ecosystem ; Pinus ; physiology ; Plants ; Poaceae ; physiology