[Determination of metabolites of styrene in urine by dispersive liquid-liquid microextraction coupled with high performance liquid chromatography].

Objective: To establish a method for the determination of mandelic acid and phenylglyoxylic acid in the urine of styrene by dispersive liquid-liquid microextraction-high coupled with high performance liquid chromatography. Methods: N-octanol was used as an extractant and ethanol was used as a dispersing agent. The phenylglycolic acid and phenylglyoxylic acid in the urine were extracted, and the upper liquid was taken after vortexing and centrifuged, and then was injected into HPLC for analysis. Results: The linear correlation coefficient of the concentration of phenylglycolic acid in the range of 0~10.0 mg/L was greater than 0.999. The detection limit of the method was 9.9 µg/L, the recovery rates were 86.1%~101.6%. The intraday RSDs of the method were 1.07%~3.76%, and the interday RSDs were 1.24%~3.33%. The linear correlation coefficient of phenylglyoxylic acid in the range of 0.0~2.0 mg/L is greater than 0.999. The detection limit of the method was 2.6 µg/L, the recovery rates were 88.8%~100.3%. The intraday RSDs of the method were 1.02%~ 3.17%, and the interday RSDs were 1.59%~2.41%. Conclusion: The method has low detection limit, high enrichment ratio and good sensitivity, and is suitable for determination of phenylglycolic acid and phenylglyoxylic acid in urine of occupational exposure to styrene.

Comparative transcriptome analysis of self-incompatible flower stigmas and self-compatible bud stigmas following self-pollination in broccoli.

DH07 is a DH line of Class I S-haplotype in broccoli (Brassica oleracea var. italica), in which stigmas of flowers show self-incompatibility (SI) and stigmas of buds show self-compatibility (SC). The molecular mechanisms that lead to stigmas at different developmental stages having different responses to self-pollination are yet unknown. In the present study, comparative transcriptome profiling of the stigmas of flowers and buds before and after self-pollination was performed by RNA-sequencing using an Illumina HiSeqTM 2000. A total of 80,102,897 reads were generated for further analysis in four libraries. Comparisons of the transcriptome profiles before and after self-pollination revealed 579 differentially expressed genes (DEGs) in the stigmas of buds (SBs); of these, 431 DEGs showed increased and 148 DEGs showed decreased transcript abundance after self-pollination in SBs. There were a total of 686 DEGs between unpollinated stigmas of flowers (SFs) and pollinated SFs, among which, 517 DEGs were up regulated and 169 DEGs were down regulated. Following the self-pollination, 379 identified DEGs were common in both SBs and SFs. It was found that ARR7-like and oxysterol-binding family protein related DEGs could play key roles in SI or SC signal transduction. The results obtained in this study would form the foundation for further studies on investigating the molecular mechanisms of SI and SC in Brassica.

In-depth analysis of internal control genes for quantitative real-time PCR in Brassica oleracea var. botrytis.

Quantitative reverse-transcription PCR (qRT-PCR) is a versatile technique for the analysis of gene expression. The selection of stable reference genes is essential for the application of this technique. Cauliflower (Brassica oleracea L. var. botrytis) is a commonly consumed vegetable that is rich in vitamin, calcium, and iron. Thus far, to our knowledge, there have been no reports on the validation of suitable reference genes for the data normalization of qRT-PCR in cauliflower. In the present study, we analyzed 12 candidate housekeeping genes in cauliflower subjected to different abiotic stresses, hormone treatment conditions, and accessions. geNorm and NormFinder algorithms were used to assess the expression stability of these genes. ACT2 and TIP41 were selected as suitable reference genes across all experimental samples in this study. When different accessions were compared, ACT2 and UNK3 were found to be the most suitable reference genes. In the hormone and abiotic stress treatments, ACT2, TIP41, and UNK2 were the most stably expressed. Our study also provided guidelines for selecting the best reference genes under various experimental conditions.

[Expression of Jagged1 mRNA in human epithelial ovarian carcinoma tissues and effect of RNA interference of Jagged1 on growth of xenograft in nude mice].

OBJECTIVE: To investigate the expression of Jagged1 in human epithelial ovarian carcinoma tissues and the effect of Jagged1 on growth of xenograft in nude mice. METHODS: (1) Forty-eight cases of ovarian cancer and 30 cases of patients with benign epithelial ovarian tumor in the Henan Province Xinxiang Central Hospital during Feb. 2011 to Mar. 2014 were enrolled in this study. The mRNA expression of Jagged1, Notch1 and the downstream target genes Hes1, Hey1 were analyzed by using realtime PCR method. (2) The ovarian cancer xenograft models in nude mice were constructed by injecting SKOV3 cells in axillary subcutaneouswere. The nude mice were randomly divided into Jagged1 interference group, blank plasmid group and control group. Each group had 10 mice. They were transfected with pcDNA3.1(+)-siRNA-Jagged1, blank plasmid pDC3.1 and phosphate buffer, respectively. The tumor volumes and tumor masses were measured 14 days after transfection and the inhibition rate was calculated. The relative mRNA expression of Jagged1, Notch1, Hes1 and Hey1 in xenograft tissues after transfection in each group was detected by using realtime PCR technique and the relative protein expression of Jagged1, Notch1, Hes1 and Hey1 in xenograft tissues was detected by utilizing western blot method. RESULTS: (1) The relative mRNA expression of Jagged1, Notch1, Hes1 and Hey1 in ovarian cancer tissues were higher than benign ovarian tumor tissues, the differences were statistically significant (P<0.01). (2) The tumor volume was (491± 68) mm(3) and tumor mass was (2.6±0.4) g in Jagged1 interference group, which were significantly lower than that in the blank plasmid group [(842±88) mm(3) and (4.4±0.8) g, respectively] and that in the control group [(851±90) mm(3) and (4.5±0.9) g, respectively; P<0.05], the tumor inhibition rate was 42.2% in Jagged1 interference group, which was significantly higher than that in the blank plasmid group and that in the control group (2.2% and 0, respectively), the differences were statistically significant (P<0.05). The relative mRNA and protein expression of Jagged1, Hes1 and Hey1 in xenograft tissues of nude micein Jagged1 interference group were lower than that in the other two groups, the differences were statistically significant (P<0.05). There were no differences of relative mRNA and protein expression of Notch1 in xenograft tissues of nude mice among the three groups (P>0.05). CONCLUSIONS: Jagged1 is highly expressed in epithelial ovarian carcinoma. Jagged1 gene interference in xenograft tumor can inhibit ovarian cancer cell growth and improve tumor suppressor rate, which probably play roles by inhibiting Notch1 signaling pathway.

Schwannomas of female genitalia from a gynaecologist's perspective: report of two cases and review of the literature.

BACKGROUND: Pelvic schwannomas are extremely rare. However, when located in the pelvic cavity, schwannomas are often encountered by a gynaecologist, not a general surgeon, and are misdiagnosed as gynaecologic masses. CASE REPORT: Here, the authors present two cases of pelvic schwannomas that were preoperatively misdiagnosed as broad ligament fibroid. One schwannoma occurred completely in the left broad ligament and was resected by laparoscopy without any complications. The other lesion was located in the retroperitoneum and had densely adhered to the surrounding tissues; this lesion was excised by laparotomy with considerable blood loss. CONCLUSIONS: Schwannomas of female genitalia are very scarce and difficulty to diagnose preoperatively. Literature review revealed 63 schwannomas arising from the female genital tract in total, 73.02% (46 cases) were located in the lower genital tract, and 26.98% (17 cases) were located in upper genital tract. The treatment modality is unique depending on the location of the tumor. Complete excision is benefical for diagnosis and treatment. The procedure can be performed safely under laparoscopy.

Clinical observational study of conformal radiotherapy combined with topotecan chemotherapy in patients with platinum-resistant recurrent ovarian cancer.

This retrospective study aimed to observe the cura-tive effect and adverse reactions of three-dimensional conformal radiotherapy combined with topotecan chemotherapy in patients with platinum-resistant recurrent epithelial ovarian carcinoma. The chemoradiotherapy group (N = 22) received 15 mv X-rays with 1.8 to 2.0 Gy/f/d radiation, 5 times per week. The total dose was 45 to 65 Gy; the median dose was 52.5 Gy. Topotecan chemotherapy (2.0 mg/m(2)) was administered after the first week of radiotherapy on days 1, 8, and 15; it was repeated every 28 days. The only che-motherapy group (N = 20) received topotecan chemotherapy (4.0 mg/m(2)) in the first week, and the dose was administered on days 1, 8, and 15; it was repeated every 28 days. The median follow-up times were 18.5 months (2 to 37.7) and 10.8 months (1.5 to 29.6) in the chemoradiotherapy and in the only chemotherapy groups, respectively. The total response rates were 42.1% (8/19) and 11.1% (2/18), respectively. The clinical benefit rates were 68.4% (13/19) and 22.2% (4/18), respectively, with significant difference (P < 0.05). The median disease progression-free periods were 9.8 and 6.6 months, respectively, with significant difference (P < 0.001). The median survival times were 19.7 and 12.5 months, respective-ly, with significant difference (P < 0.05). The degrees of digestive tract reaction rates were 26.3% (5/19) and 16.7% (3/18), whereas the hematology toxicity rates were 21.1% (4/19) and 22.2% (4/18), respectively, with no significant difference (P > 0.05). As three-dimensional conformal radiotherapy combined with topotecan che-motherapy had good curative effect on platinum-resistant recurrent epithelial ovarian cancer, with mild adverse reactions, this tech-nique can be used as a remedial measure.

Which is the appropriate surgical procedure for Stage I endometrial carcinoma?

OBJECTIVE: To study the appropriate surgical procedure for Stage I endometrial carcinoma (EC), the clinical and pathological features and prognosis factors, as well as types were analyzed retrospectively. MATERIALS AND METHODS: This is a retrospective study of 277 patients with early-stage EC in clinical Stages I that received surgery between January 2000 and March 2008. The appropriate surgical procedures were divided into three types (procedure I-III: hysterectomy with or without ovary preservation, subradical hysterectomy plus pelvic lymph node biopsy, and radical hysterectomy pelvic plus lymphadenectomy) according to the clinical stage. RESULTS: Tumor invasion of the cervix and deep muscularis as well as the parametrium, EC Stage Ib, grade 3 and ascites had carcinoma cells, were high-risk factors of EC metastasis to the retroperitoneum (p < 0.05). The ovarian preservation of EC Stage Ia had no effect on overall survival. The three types of procedure for the EC Stage Ia were not correlated significantly to the three-year and five-year survival rates. The three-year and five-year survival rates of three surgical procedures for the EC Stage Ib were significantly correlated. The survival rates of surgical procedures II and III were significantly higher than that of procedure I (P < 0.05). CONCLUSION: Subradical hysterectomy plus pelvic lymph node biopsy was recommended for EC Stage 1b with high-risk factors. There was no evidence of benefit in terms of overall or recurrence-free survival for radical hysterectomy plus pelvic lymphadenectomy in women with Stage I EC.