Rapid screening of glycosyltransferases in plants using a linear DNA expression template based cell-free transcription-translation system.

Plants have an extensively large number of enzymes including glycosyltransferases that are important in the biosynthesis of natural products. However, it is time-consuming and challenging to study these enzymes and only a small percentage of them have been well-characterized. Here, we report a rapid method to screen plant glycosyltransferases using a linear DNA expression template (LET) based cell-free transcription-translation system (TX-TL). As a proof of concept, we amplified and tested glycosyltransferases from Arabidopsis thaliana and showed that the catalytic activity results of these glycosyltransferases from LET-based-TX-TL were consistent with previous studies. We then chose a local medicinal plant Anoectochilus roxburghii, acquired its transcriptome sequences, and applied this method to study its glycosyltransferases. We rapidly expressed all the putative UDP-glucose glycosyltransferases using LET-based-TX-TL and discovered 6 unreported active glycosyltransferases which can catalyze the glycosylation of quercetin into isoquercitrin. Thus, LET-based-TX-TL was shown to be a powerful tool for researchers to rapidly screen plant glycosyltransferases for the first time.

Quantitative Analysis of Eight Triterpenoids and Two Sesquiterpenoids in Rhizoma Alismatis by Using UPLC-ESI/APCI-MS/MS and Its Application to Optimisation of Best Harvest Time and Crude Processing Temperature.

Rhizoma Alismatis (RA), widely known as "Ze-Xie" in China, is the tuber of Alisma orientale (Sam.) Juzep (Alismaceae), a Chinese herbal medicine that has been used to treat hyperlipidemia, diabetes, hypertension, dysuria, and inflammation. In this study, a sensitive and reliable method based on an ultra-performance liquid chromatography (UPLC) couple with two ionisation modes, including electrospray ionisation (ESI) and atmospheric pressure chemical ionisation (APCI) tandem mass spectrometry (MS), namely, UPLC-ESI/APCI-MS/MS was developed and validated to simultaneously determine 8 triterpenoids (ESI mode) and 2 sesquiterpenoids (APCI mode) in RA. Ten marker compounds were analysed with a Waters' CORTECS UPLC C18 column (200 mm × 2.1 m, 1.6 µm) and gradient elution with water (contained 0.1% formic) and acetonitrile within 7 min. The established method was validated for linearity, intra- and interday precisions, accuracy, recovery, and stability. The calibration curve for 10 marker compounds showed good linear regression (r > 0.9971). The limits of detection and quantification for analytes were 0.14-1.67 ng/mL and 0.44-5.65 ng/mL, respectively. The relative standard deviations (RSD, %) and accuracy (RE, %) of intra- and interday precisions were less than 3.83% and 1.21% and 3.22% and 1.46%, repeatability and stability for real samples were less than 2.78% and 3.19%, respectively. All recoveries of the 10 marker compounds ranged from 97.24% to 102.49% with RSDs less than 4.05%. The developed method efficiently determined the 10 marker compounds in RA and was subsequently applied to optimise harvest time and crude processing temperature. The result indicated the 90% wilted phase and 70°C (or lower) may be the best harvest time and the processing temperature of RA.

[Study on UHPLC fingerprint and determination of eight phenolic components of Tetrastigma hemsleyanum leaves].

A novel method combining ultra-high performance liquid chromatography (UHPLC) fingerprint and simultaneous quantitative analysis of eight phenolic components was developed and validated for quality evaluation of Tetrastigma hemsleyanum leaves. For fingerprint analysis, 15 peaks were selected as the common peaks to evaluate the similarities among 41 batches of T. hemsleyanum leaves collected from different regions. Additionally, simultaneous quantification of eight markers, including neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isoorientin, orientin, vitexin-2-O-rhamnoside,vitexin and isovitexin, was performed and the obtained data demonstrated that our method has achieved desired linearity, precision and accuracy. Clustering statistical analysis was further application in T. hemsleyanum leaves from different regions. The results indicated that new approach conbine ultra-high performance liquid chromatography (UHPLC) fingerprint and simultaneous quantitative analysis of eight phenolic components was applicable in quality control of T. hemsleyanum leaves.

Axillary Metastasis as the First Manifestation of Occult Breast Cancer in a Male Patient.

SUMMARY: BACKGROUND: The aim of this study was to investigate the clinicopathologic features of male breast cancer. CASE REPORT: We present the clinicopathologic data of a 72-year-old male patient with occult breast cancer, who was diagnosed and underwent surgery in our hospital. The diagnosis was confirmed by histological examination, and the patient underwent modified radical mastectomy and axillary dissection. The histological examination showed no tumor foci in the resected breast tissue, but 2 of 15 dissected axillary lymph nodes were invaded by infiltrating ductal carcinoma. Immunohistochemistry staining was negative for both estrogen and progesterone receptors, but showed expression of p53 protein (+++), proliferating cell nuclear antigen (PCNA) (+++), Bcl-2 on-coprotein (+++), nm23 protein (++), multidrug resistance protein (MRP) (++), and human epidermal receptor (HER-2) oncoprotein (+++). 24 months after being diagnosed, the patient is alive without any residual or metastatic disease. CONCLUSIONS: Breast cancer is very rare in men, and the occurrence of occult breast cancer is even less common. Axillary metastases can present as the first manifestation of breast cancer in a male.

Axillary metastasis as the first manifestation of male breast cancer: a case report.

BACKGROUND: Breast cancer is very rare in men, and the occurrence of occult breast cancer which present axillary metastasis as the first manifestation is even rarer in men. CASE PRESENTATION: We report a 72-year-old male Han-Chinese patient who presented axillary metastasis as the first manifestation of breast cancer and got correctly diagnoses by histological examination. He underwent modified radical mastectomy and axillary dissection on 11 Apr 2006. The histopathologic examination showed that no tumor focus was found in his breast tissue, but two out of fifteen of axillary lymph nodes were invaded by infiltrating ductal carcinoma. The IHC stain showed that estrogen receptor (ER) and progestin receptor (PR) were negative, Human epidermal receptor (HER-2) oncoprotein (+++), P53 protein expressed (+++), Bcl-2 oncoprotein (+++), nm23 protein (++), proliferating cell nuclear antigen (PCNA) (+++) and multidrug-resistance protein (MRP) (++). After operation, he did not receive endocrine therapy, chemotherapy and radiotherapy because of his senility. He is alive without any residual or metastasis disease 29 months after being diagnosed. CONCLUSION: This is the first case in our hospital that presents axillary metastases as the first manifestation of male breast cancer.

Sclerosing mesenteritis as a rare cause of abdominal pain and intraabdominal mass: a cases report and review of the literature.

Sclerosing mesenteritis is a rare, benign, and chronic fibrosing inflammation disease with unknown etiology that affects the mesentery of small bowel and colon. The disease has two well-established histological types: the acute or subacute form known as mesenteric panniculitis and the chronic form known as retractile or sclerosing mesenteritis. Because the sclerosing mesenteritis is lack of special clinical manifestation and typical signs, so the patients are very easy to be misdiagnosed. The correct diagnosis of sclerosing mesenteritis depends on pathological examination and exploratory laparotomy. We report a case of sclerosing mesenteritis in a 52-year-old male who presented with chronic abdominal pain and intraabdominal mass. This patient had a long-term and heavy drinking history. He was misdiagnosed as celiac teratoma by CT examination and then underwent an exploratory laparotomy at March 2 2004. A mass, its diameter being about 5 cm, was detected in mesentery of distal ileum. Although a few small intestines tightly adhered on the mass, the involved intestine had no obstruction. The intraoperative biopsy indicated that it was an inflammatory mass. The mass and adhered intestines were removed. He was diagnosed with sclerosing mesenteritis by histopathological examination of paraffin section. After operation, this patient went well and lives without recrudescence at the time we wrote this paper.