Low Concentration of Wenyang Tonglin Decoction Promotes Conjugation and Transfer of Drug-Resistant Plasmids among Heterologous Strains.

OBJECTIVE: To investigate the effect of low concentration of Wenyang Tonglin Decoction (WTD) on the binding conditions of R45 plasmid conjugative transfer under liquid phase conjugation and its mechanism. METHODS: Escherichia coli CP9 (R45) and Staphylococcus aureus RN450RF were cultured in medium containing WTD, and their minimum inhibitory concentration (MIC) values were obtained. Using promoter fusion technology, E. coli CP9 (R45) containing a promoter fusion was obtained. ß-Galactosidase activity of TrfAp and TrbBp was tested, and the mRNA expression of regulatory factors (TrbA, KorA, and KorB) was detected by real-time fluorescent quantitative polymerase chain reaction. RESULTS: The MIC of E. coli CP9 (R45) was 400 g/L and that of S. aureus RN450RF was 200 g/L. When the drug concentration in the culture medium was 200 g/L, the highest number of conjugants was (3.47 ±0.20) × 107 CFU/mL At 90 h of conjugation, the maximum number of conjugants was (1.15 ±0.06) × 108 CFU/mL When the initial bacterial concentration was 108 CFU/mL, the maximum number of conjugants was (3.47 ± 0.20) × 107 CFU/mL. When the drug concentration was 200 g/L, the ß-galactosidase activity of TrfAp and TrbBp significantly increased; the relative quantification of TrbA, KorA and KorB were significantly inhibited. CONCLUSION: Low concentration of WTD promoted the development of bacterial resistance by affecting promoters and inhibiting the expression of regulatory factors.

[Clinical observation of acupuncture combined with moxibustion on college students with depression].

OBJECTIVE: To observe the clinical efficacy of acupuncture combined with moxibustion with seed-sized moxa cone in the treatment of depression in college students, and explore its effect on serum 5-hydroxytryptamine (5-HT). METHODS: Sixty undergraduates with depression were divided into acupuncture-moxibustion group and medication group according to the random number table, with 30 patients in each group. The patients in acupuncture-moxibustion group received acupuncture and moxibustion with seed-sized moxa cone at acupoints on Shaoyang meridian according to the method of "rotating the pivot and regulating the qi", one time every other day. The patients in medication group received oral administration of fluoxetine hydrochloride capsule 20 mg once a day, and both groups were treated continuously for 8 weeks. The scores of Hamilton depression (HAMD-17) scale, self-rating depression scale (SDS) and serum 5-HT content before and after treatment were compared between the two groups. RESULTS: Compared with before treatment, the HAMD-17 and SDS scores of the acupuncture-moxibustion group began to decrease after 2 weeks of treatment (P<0.05); while the HAMD-17 and SDS scores of the the medication group began to decrease after 4 weeks of treatment (P<0.05). After 2, 4 and 8 weeks of treatment, the HAMD-17 and SDS scores of the acupuncture-moxibustion group decreased more significantly than the medication group (P<0.05). The 5-HT contents of the two groups were significantly higher than those before treatment (P<0.05)，and there was no significant difference in serum 5-HT content between the two groups (P>0.05). The adverse reaction score of the acupuncture-moxibustion group was lower than that of the medication group (P<0.05). The total effective rate of the acupuncture-moxibustion group was 92.86%（26/28）, better than the medication group 81.48% (22/27,P<0.05). CONCLUSION: Acupuncture combined with seed-sized moxa cone moxibustion is more effective than oral administration of fluoxetine hydrochloride in treating college students' depression, and acupuncture combined with moxibustion has a faster onset and fewer adverse reactions in the treatment of college students' depression.

[Effects of mild moxibustion on Beclin-1/Bcl-2 expression in spinal cord of rats with cervical spondylotic radiculopathy].

OBJECTIVE: To observe the effects of mild moxibustion on the expression of autophagy and apoptosis factors Beclin-1, Bcl-2 mRNA and protein in spinal cord (including nerve root tissues) of cervical spondylotic radiculopathy (CSR) rats, so as to explore the analgesic mechanism of mild moxibustion at "Dazhui" (GV14) on CSR. METHODS: SD rats were randomly divided into blank group, model group, mild moxibustion group and mild moxibustion+3-methyladenine(3-MA) group, with 10 rats in each group. CSR model was established by inserting the wire into the cervical nerve root. The rats in the blank group were only fed normally without any intervention.The rats in the mild moxibustion group and mild moxibustion+3-MA group were given mild moxibustion at GV14 for 10 min each time,and intraperitoneal injection of 1 mL 0.9% normal saline and 1 mL 3-MA(15 mg/kg)separately. Rats in the model group were given 0.9% normal saline every day. All the three interventions were started from the 3rd day after modeling for 7 days. The rat's behavioral reaction of gait was scored and the pain threshold of rat was measured with a pain analyzer; the expressions of Beclin-1 and Bcl-2 mRNA and protein in the spinal cord (including nerve root) were detected by fluorescence quantitative PCR and immunohistochemistry, separately. The autophagosome and ultrastructure of the spinal nerve root tissue were observed by transmission electron microscope. RESULTS: After modeling, the gait score was significantly increased (P<0.01) and the pain threshold significantly decreased (P<0.01) in the model group in comparison with the blank group. There was no statistical difference in Beclin-1 and Bcl-2 mRNA and protein expression between the blank and the model groups. After intervention, compared with the model group, the gait scores were significantly reduced (P<0.01), the pain threshold and the expressions of Beclin-1 and Bcl-2 mRNA and protein were significantly increased (P<0.01，P<0.05) in the mild moxibustion and mild moxibustion+3-MA groups. The improvement of the above indicators as more significant in the mild moxibustion group than that in the mild moxibustion+3-MA group (P<0.05). After modeling, the organelles in the spinal nerve root tissue cells of the model group were damaged and there were a small amount of autophagosomes. Compared with the model group, the ultrastructure of the spinal nerve root tissue cells in the mild moxibustion group were relatively complete, and the number of autophagosomes increased. CONCLUSION: Mild moxibustion at GV14 has a good analgesic effect on CSR rats, which may be related to its effects in up-regulation of Beclin-1 and Bcl-2 expressions and activation of autophagy and inhibition of apoptosis.

Poly[bis-(µ(4)-acetato-κO:O:O':O')bis-(µ(3)-acetato-κO:O:O)(µ(2)-acetato-κO:O')(µ(2)-acetic acid-κO:O')di-µ-aqua-copper(II)tris-odium].

In the title compound, [CuNa(3)(CH(3)CO(2))(5)(CH(3)COOH)(H(2)O)(2)](n), the Cu(II) atom lies on an inversion center and is coordinated by four O atoms from four acetate ligands, leading to a square-planar geometry. One Na(I) atom, lying on an inversion center, is coordinated by four O atoms from four acetate ligands and two bridging water mol-ecules in a distorted octa-hedral geometry. The other Na(I) atom is coordinated by five O atoms from five acetate ligands and a bridging water mol-ecule. A hy-droxy H atom lies on a twofold rotation axis and is shared by two acetate ligands. The crystal packing exhibits a polymeric layer parallel to (100), which is further stablized by intra-layer O-H⋯O hydrogen bonds. The layers are linked by inter-layer O-H⋯O hydrogen bonds.

Complete profiling and characterization of in vitro nefazodone metabolites using two different tandem mass spectrometric platforms.

This paper describes the complete profiling and characterization of in vitro metabolites of the antidepressant agent nefazodone (NEF) generated by human liver microsome (HLM). Two new metabolic pathways (biotransformation) for NEF have been discovered by the characterization of three new metabolites, including two new metabolites (M24, M25) formed due to the N-dealkylation reaction that occurred between the triazolone and propyl units, and one new metabolite (M26) formed due to the O-dearylation reaction that occurred on the phenoxyethyl unit. These metabolites were initially detected by a 4000 Q-Trap instrument and then confirmed by exact mass measurement using an LTQ-Orbitrap. Both instruments proved to be capable of providing complete in vitro metabolite information in a single liquid chromatography/tandem mass spectrometry (LC/MS/MS) analysis, although each had its advantages and disadvantages. One noticeable disadvantage of the 4000 Q-Trap was the reduced quality of isotopic pattern in the enhanced mass scan (EMS) spectrum when it was used as survey scan to trigger multiple dependent product ion scans. The problem was especially exacerbated for minor metabolites with low signal intensity. On the other hand, the LTQ-Orbitrap maintained excellent isotopic pattern when used as a full scan survey scan. Twenty-six metabolites were detected and identified. The formation of these new metabolites was also confirmed by analyzing duplicate incubations at different time points.

Direct injection of solid-phase extraction eluents onto silica columns for the analysis of polar compounds isoniazid and cetirizine in plasma using hydrophilic interaction chromatography with tandem mass spectrometry.

Isoniazid and cetirizine do not retain well on reversed-phase columns due to their high polarity. Silica columns, when operated under hydrophilic interaction conditions, do provide excellent retention of these compounds. We have developed simple and proof of concept analytical methods for the analysis of isoniazid and cetirizine in animal and human plasma, respectively. Both methods employed the approach of direct injection of solid-phase extraction (SPE) organic eluents onto silica columns for analysis, thus eliminating evaporation and reconstitution steps that are typically needed for reversed-phase liquid chromatographic analysis. Isoniazid was extracted from animal plasma samples using a Waters Oasis HLB 96-well plate and then eluted with acetonitrile, while cetirizine was extracted from human plasma with a Waters MCX mu-Elute plate and then eluted with acetonitrile containing 5% concentrated ammonium hydroxide. The direct injection of the SPE eluent onto the analytical column was necessary since significant loss of isoniazid was found during the evaporation and reconstitution steps. The method for isoniazid also enabled ultra-fast analysis due to the relatively low back-pressure exhibited by silica columns even under high flow conditions. Both methods show good linearity, accuracy and precision covering the range of 10-2000 ng/mL of isoniazid, and 1-1000 ng/mL of cetirizine in plasma. Substantial time savings were realized as a result of both the elimination of the evaporation and reconstitution steps and the fast chromatographic analysis.