To synthesize aristolochic acid (AA)-2'-deoxyguanosine 5'-monophosphate (dGp) adducts in vitro and develop a novel method for the characterization of the adducts using multiple mass spectrometric techniques. AA was incubated with dGp in vitro using either enzymatic activation (by xanthine oxidase) or chemical activation (by zinc) to synthesize AA-dGp adducts, and the reaction conditions were optimized. Crude extracts were analyzed by techniques of liquid chromatography-electrospray ionization/tandem mass spectrometry (LC-MS/MS) and high accuracy mass data and isotope pattern of super high resolution Fourier transform-ion cyclotron resonance mass spectrometry (FT-ICRMS). The quasi-molecular ion peaks of the AA-dGp adducts were obtained in the negative ion mode. Analysis by electrospray ionization/tandem mass spectrometry (ESI-MS/MS) provided useful structural information about AA-dGp adducts. AA can bind covalently to the exocyclic amino group of deoxyguanosine to form AA-dGp adducts. MS analysis is a powerful tool to detect and identify AA-dGp adducts simply, rapidly and accurately.
Aristolochic Acids ; chemical synthesis ; chemistry ; Chromatography, High Pressure Liquid ; methods ; DNA ; chemistry ; metabolism ; DNA Adducts ; chemical synthesis ; Deoxyguanosine ; chemistry ; Tandem Mass Spectrometry ; methods

OBJECTIVETo optimize the best acupuncture therapy in the treatment of lumbar disc herniation (LDH) and to explore its mechanism in terms of immunology.

METHODSOne hundred and eleven cases were randomized into an acupuncture-moxibustion group, an elongated needle group and a herb-partitioned moxibustion group, 37 cases in each group. In acupuncture-moxibustion group, penetration needling with elongated needle was applied at Dachangshu (BL 25), Guanyuanshu (BL 26) and Huantiao (GB 30), which was followed with herb-partitioned moxibustion at Shenque (CV 8) and Yaoyangguan (GV 3), etc, once a day. Elongated needle and herb-partitioned moxibustion were applied in the elongated needle group and herb-partitioned moxibustion group respectively. The efficacy of each group was observed after 21 treatments and the value of serum immunoglobulin was measured and analyzed before and after treatment.

RESULTSThe effective rate was 97.3% (36/37) in acupuncture-moxibustion group, which was superior to 75.7% (28/37) in the elongated needle group and 73.0% (27/37) in the herb-partitioned moxibustion group (both P<0.05). The visual analogue scale (VAS), straight-leg raising range and motion range of the lumbar vertebra in three groups were all improved after treatment (all P<0.05), which was more significant in the acupuncture-moxibustion group (all P<0.05). The difference of value of serum immunoglobulin before and after treatment was significant in the acupuncture-moxibustion group (P<0.05), which was not in the elongated needle group and the herb-partitioned moxibustion group (both P>0.05).

CONCLUSIONThe therapy of elongated needle combined with herb-partitioned moxibustion achieves superior efficacy in the treatment of LDH as compared with elongated needle or herb-partitioned moxibustion therapy, and it can regulate the humoral immunity system of patients.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; Adult ; Female ; Humans ; Intervertebral Disc Displacement ; therapy ; Male ; Middle Aged ; Moxibustion ; Treatment Outcome ; Young Adult

RecQ5 in mammalian cells has been suggested to suppress inappropriate homologous recombination. However, the specific pathway(s) in which it is involved and the underlining mechanism(s) remain poorly understood. We took advantage of genetic tools in Drosophila to investigate how Drosophila RecQ5 (dRecQ5) functions in vivo in homologous recombination-mediated double strand break (DSB) repair. We generated null alleles of dRecQ5 using the targeted recombination technique. The mutant animals are homozygous viable, but with growth retardation during development. The mutants are sensitive to both exogenous DSB-inducing treatment, such as gamma-irradiation, and endogenously induced double strand breaks (DSBs) by I-Sce I endonuclease. In the absence of dRecQ5, single strand annealing (SSA)-mediated DSB repair is compromised with compensatory increases in either inter-homologous gene conversion, or non-homologous end joining (NHEJ) when inter-chromosomal homologous sequence is unavailable. Loss of function of dRecQ5 also leads to genome instability in loss of heterozygosity (LOH) assays. Together, our data demonstrate that dRecQ5 functions in SSA-mediated DSB repair to achieve its full efficiency and in suppression of LOH in Drosophila.
Animals ; DNA Repair ; genetics ; DNA, Single-Stranded ; genetics ; Drosophila Proteins ; genetics ; metabolism ; Drosophila melanogaster ; genetics ; metabolism ; Loss of Heterozygosity ; genetics ; RecQ Helicases ; genetics ; metabolism

Objective:To study the effect of physical therapy and heel sound feedback on lower limbs motor function, mobility and activities of daily living (ADL) for stroke patients based on International Classification of Functioning, Disability and Health (ICF) core set. Methods:From April, 2018 to May, 2020, 113 stroke patients with motor dysfunction were divided into ischemia group (n = 67) and hemorrhagia group (n = 46) according to the cause of stroke. They received physical therapy for lower limbs and heel sound feedback for eight weeks, and assessed with ICF core set for stroke-gait, Fugl-Meyer Assessment-Lower Extremities (FMA-LE), Timed 'Up and Go' Test (TUGT), and modified Barthel index (MBI) before and after intervention. Results:The main effect of time was significant for qualifiers of ICF core set for stroke-gait, the scores of FMA-LE and MBI, and TUGT time (F > 100.59, P < 0.001), and it improved time by time as Post Hoc test. The main effect of groups was not significant (F < 2.29, P > 0.05), nor as Post Hoc test. The interactive effect between time and groups was significant for TUGT time (F = 6.45, P < 0.01), perhaps improved more in the hemorrhagia group, however, the interactive effect was not significant for the others. Conclusion:Physical therapy and heel sound feedback can improve motor function of lower limb, mobility and ADL for stroke patients.