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A liquid chromatography tandem mass spectroscopy method was employed to determine free and protein-bound Nε-(carboxymethyl)lysine (CML) in seven categories of dairy products (liquid milk n = 28, milk powder n = 26, condensed milk n = 6, milk fats n = 4, cheese n = 9, ice cream n = 6, and whey protein n = 3) commonly consumed in China. Recovery of the analyte based on the isotopic internal standard correction ranged from 104 to 115%, with relative standard deviations of less than 5.13%. The limit of quantification was 0.1 μg/kg. An extremely significant difference was found in the protein-bound CML content between pasteurized milk and sterilized milk (P < 0.01), there was a significant difference in the free CML content between pasteurized milk and sterilized milk (P < 0.05). The average content of free CML (14.02 μg/kg) and protein-bound CML (5.56 mg/kg) in pasteurized milk were higher than that (8.23 μg/kg, 2.58 mg/kg) in sterilized milk. The mean value of CML in milk powder were 1.60 mg/kg sample. The highest levels of free CML (32.71 ± 3.67 μg/kg sample) and protein-bound CML (134.28 ± 11.77 mg/kg sample) were detected in whey protein powder. [ABSTRACT FROM AUTHOR]

Objective: To study the antagonistic effect of oral contraceptives (OCs) on the tripterygium wilfordii polyglycoside (TWP)-induced gonad toxicity in the female SD rats. Methods: 40 female SD rats were divided randomly into four groups: the TWP group, the OCs group, the TWP with OCs group, and the blank control group. Test of vaginal exfoliated cell smears was done everyday after 10 weeks' treatment. Rats were sacrificed after stopping treatment 3 weeks. Serum follicle stimulating hormone (FSH),luteinizing hormone (LH), estradiol (E2) were measured by ABC-ELISA. The weights of uterus and ovaries, and the number of follicles, were compared among groups. The histomorphology of uterus and ovary was observed by microscope. Results: The oestrous cycle in the TWP group was incomplete or extended; serum E2 level [(40.417 4±4.937 2) ng/L] was significantly lower than those in other groups; while FSH level[( 1.063 4±0.153 9) IU/L] and LH levels [(5.946 7± 1.036 0) IU/L] were significantly higher than those in other groups (P<0.01). The weight indexes of uterus (0.048 7%± 0.009 7%) and ovaries (0.147 0% ±0.004 7%), and the thickness of endometrium [(57.20 ±1.81) µm] and myometrium [(24.40±1.84) µm],were significantly lower than those in other groups (P<0.01). The numbers of growing follicle (22.50±2.55) and graaffian follicle (2.00±1.89) in the TWP group were significantly less than those in the other groups (P<0.05),while there was not significant difference in the number of primordial follicles (36.50± 2.64) in the TWP group when compared with other groups (P>0.05). Conclusions ;Diane-35 can decrease the TWP-induced gonad damage in female SD rats, which plays a potential role in protecting ovarian function. [ABSTRACT FROM AUTHOR]

Objective: To validate the therapeutic efficacy of paroxetine in the treatment of premature ejaculation (PE)., Methods: Eighty PE patients up to the inclusion criteria were equally randomized to an experimental and a control group. We observed all the patients for 4 weeks and recorded the baseline data on intravaginal ejaculatory latency time (IELT) and sexual satisfaction scores, followed by oral medication of paroxetine at 20 mg/d for the patients in the experimental group and placebo for the controls. Thirty days after the treatment, we again recorded IELT and sexual satisfaction scores of the patients., Results: After the treatment, the experimental group showed significantly prolonged IELT ([5.75 +/- 1.24] min) and increased sexual satisfaction score (6.4 +/- 1.2) as compared with the baseline data ([0.89 +/- 0.21] min and [2.7 +/- 0.9]) (P < 0.01). The control group exhibited no significant differences before and after the medication either in the mean IELT or in sexual satisfaction scores ([1.06 +/- 0.28] min vs [0.97 +/- 0.18] min and 3.6 +/- 1.3 vs 3.1 +/- 1.1, P > 0.05)., Conclusion: Oral medication of paroxetine at 20 mg/d for 30 days could improve IELT and sexual satisfaction in PE patients.

Objective: To investigate the mechanisms of sodium selenite induced DNA damage in HepG2 cells., Methods: HepG2 cells were treated with the designed concentrations of sodium selenite and the selenite (10 micromol/L) added simultaneously with GSH (10 mmol) and NAC (5 mmol). Then the cell viability was detected by MTT, and the flurescent intensity of reactive oxygen species (ROS) was determined by flow cytometry, and DNA damage was detected by commet assay., Results: The level of ROS was increased after HepG2 was treated with 5, 10, 20 micromol/L sodium selenite for one hour, and the cell viability was decreased after 12 hours, and the DNA damage was enhanced. Compared with the control group, the difference was statistically significant (P < 0.05) . GSH and NAC effectively inhibited the ROS increased and cell viability decreased and DNA damage weakened., Conclusion: ROS may be the important reason that sodium selenite induced HepG2 cells DNA damage.

Objective: To investigate and compare the expression of Toll-like receptors (TLRs) in human bladder cells under the stimulus of Escherichia coli (E. coli) versus that under the stimulus of Bacillus Calmette-Guérin (BCG)., Methods: Human bladder cancer cell line T24 was cultured for 1 hour under the stimuli of various doses (bacillus-cell ratio) of E. coli and BCG respectively. The levels of expression of TLR-2 and TLR-4 mRNA in T24 cells were assessed by RT-PCR method., Results: By comparison with control, there was no difference observed on the expression of either TLR-2 or TLR-4 in T24 cells under the stimulus of E. coli. The expression level of either TLR-2 or TLR-4 was increased under the stimulus of BCG in a dose-dependent manner. The effects reached the highest level when the dose of BCG was 10 bacilli per cell., Conclusion: There exist different expression patterns of TLRs in bladder transitional cells under the different stimuli of E. coli and BCG respectively.

Objective: To investigate the effect of bacillus Calmette-Guerin (BCG) on Toll-like receptors (TLRs) expression and cytokine production in human bladder cancer cell line T24., Methods: Human transitional carcinoma cell line T24 was cultured in RPMI1640 medium with 10% FBS, BCG was added into the cell culture with various doses in bacteria-cell ratio. After T24 cells were stimulated by BCG for 1 hour, total RNA of cells was extracted. RT-PCR procedure was conducted with the primer of TLR-2 and TLR-4, and the products were analyzed with agarose gels electrophoresis. Then the expression of TLR-2 and TLR-4 in T24 cells was assessed by the analyzing system of gel imaging. After the stimulation culture for 12 hours, the supernatant of cells was collected. The levels of IL-4 and IL-12 in each sample were assayed by ELISA method., Results: The expression level of either TLR-2 or TLR-4 was increased by the stimulation of BCG in a dose-dependent manner, the effects reached the maximal level at the dose of BCG as 10 bacilli per cell. The production of IL-12 in T24 cells was also increased gradually by the stimulation of BCG in a dose-dependent manner, and the dose of BCG obtained maximal effect was 10 bacilli per cell, which is coincident with the results observed in the expression of TLRs. There was no difference showed on the production of IL-4 between T24 cells stimulated with BCG and control., Conclusions: The stimulation of BCG not only up-regulated the expression of TLR-2 and TLR-4, but also increased the production of IL-12 in bladder cancer T24 cell line. The expression of TLRs and the production of cytokine in bladder cancer cells may be related to the BCG-induced immunol response to human bladder cancer.