[Establishment and characterization of patient derived xenograft model of malignant peritoneal mesothelioma in nude mice].

Objective: To establish patient derived xenograft (PDX) model of malignant peritoneal mesothelioma (MPM), and to identify the key characteristics of tumor biology of the model, so as to provide an experiment platform for studying the pathologic mechanisms and new therapeutic strategies for MPM. Methods: Surgically excised MPM tumor tissues were inoculated subcutaneously in BALB/c-nu/nu mice for 3 stable passages. In the 4th passage, the subcutaneous tumors were harvested under aseptic conditions, cleaned and made into MPM tumor cell homogenate. Four nude mice (two males and two females) were selected and one male and one female nude mouse were inoculated in the abdominal cavity at the dose of 100 µL, others were inoculated at a dose of 200 µL. The PDX model of MPM was established. The changes of body mass in nude mice were measured regularly, the extent of abdominal and pelvic tumors was judged by experimental peritoneal cancer index (ePCI) score, and the pathologic characteristics of tumors were analyzed. Results: The subcutaneous and abdominal animal models of MPM were successfully established. The subcutaneous tumor model grew into tumor on the 20th day, followed by a slow growth stage between the 20th and 29th day, then a rapid growth stage between the 30th and 57th day. According to the dose of tumor cells (100, 200 µL) and timing (14th and 69th days after grafting), the abdominal tumor model successfully simulated the early and late clinical stages of MPM. The HE staining results of the MPM nude mice model showed that the tumor was epithelial mesothelioma and invaded most of the organs, including liver, spleen, pancreas, mesentery. Immunohistochemical staining for calretinin, cytokeratin 5/6, WT1 and Ki-67 were positive. Whole-genome exon sequencing identified 26 and 36 high frequency gene mutations in tumors derived from the PDX model and clinical sample from patients, including 21 common gene mutations. Conclusions: The PDX model of MPM is established. The model is characterized by highly malignant tumor with rapid growth and high invasiveness.

[Recurrence pattern and influencing factors of intensity-modulated radiotherapy for cervical cancer].

With the rapid development of precise radiotherapy technology, the use of intensity-modulated radiation therapy (IMRT) in gynecological malignant tumor has been gradually increased. IMRT could result in a good local control rate as well as superior clinical outcome in patients with cervical cancer after radical radiotherapy. The recurrence pattern of radiotherapy includes in-field recurrence, out-of-field and combined failure. The recurrence pattern of radical IMRT for cervical cancer is mainly out-of-field recurrence. Out-of-field recurrence may be related to inadequate imaging evaluation before treatment. In-field recurrence may be related to insufficient radiation dose in target area and differences of tumor sensitivity.

[Effect of iron overload on the level of carbohydrate antigen 199 in type 2 diabetes mellitus].

Objective: To investigate the effect of iron overload on tumor marker CA199 in patients with type 2 diabetes mellitus (T2DM). Methods: Between January 2017 and June 2018, three hundred and twenty-three hospitalized patients with T2DM in Department of Endocrinology, Tianjin Fourth Central Hospital were selected as diabetes group and another 100 healthy persons as control group. All the participants with major diseases and factors affecting serum ferritin and CA199 level were excluded. Serum ferritin and CA199 levels were measured by chemiluminescence assay. Patients with a ferritin level above the normal range were defined as iron overload group. According to serum ferritin level, 178 patients were divided into iron overload group and 145 into normal ferritin group in the diabetes group. The levels of CA199 were compared between iron overload group and normal ferritin group, and the correlation was analyzed. Results: The levels of serum ferritin [407.1 (346.7) µg/L vs 266.6 (201.1) µg/L in males, 184.7 (133.0) µg/L vs 77.4 (64.0) µg/L in females,both P<0.001] and CA199 [18.2 (19.3) µg/L vs 9.3 (9.7) µg/L, P<0.001] in diabetes group were significantly higher than those in control group. Serum CA199 levels in iron overload group were significantly higher than those in normal ferritin group (20.7 µg/L vs 15.0 µg/L, P<0.001; above intermediate value percent:53.9% vs 35.8%,P=0.001). CA199 was positively correlated with glycosylated hemoglobin (HbA1c) (r=0.280, P<0.001) and iron overload (r=0.210, P<0.001). Multiple linear regression analysis showed that iron overload (ß'=0.203, P=0.002) and HbA1c (ß(')'=0.202, P=0.001) had a significant effect on CA199 level. Conclusion: In T2DM patients, iron overload is an important factor affecting serum CA199 level.

Association between aldehyde dehydrogenase 2 (ALDH2) Glu504Lys polymorphism and susceptibility to colorectal cancer: a meta-analysis.

Numerous studies have evaluated the association between Glu504Lys polymorphism in the aldehyde dehydrogenase 2 (ALDH2) gene and colorectal cancer (CRC) risk. However, the specific association remains controversial. To assess the relationship between the ALDH2 Glu504Lys polymorphism and CRC, we conducted a comprehensive meta-analysis of five case-control studies comprising 1664 patients with CRC and 2777 controls. The results of this meta-analysis showed that the ALDH2 Glu504Lys polymorphism was associated with a significantly reduced risk of CRC [Lys/Lys vs Glu/Glu: odds ratio (OR) = 0.95, 95% confidence interval (CI) = 0.58-1.54; Glu/Lys vs Glu/Glu: OR = 0.85, 95%CI = 0.75-0.97; dominant model: OR = 0.86, 95%CI = 0.76-0.98; recessive model: OR = 1.00, 95%CI = 0.62-1.61]. No significant heterogeneity or publication bias was observed in our meta-analysis. Based on the statistical data, our meta-analysis indicates that the ALDH2 Glu504Lys polymorphism is associated with reduced risk of developing CRC.

[Expression and clinical significance of CD147 in parathyroid carcinoma].

OBJECTIVE: To study the expression and clinical significance of CD147 in the patients of parathyroid carcinoma. METHODS: Fourteen cases of parathyroid carcinoma encountered during the period from 2012 to 2015 were enrolled. Thirty three cases of parathyroid adenoma encountered during the same period were enrolled. The expression of CD147 in parathyroid carcinoma and parathyroid adenoma was studied by means of immunohistochemistry (EnVision method). RESULTS: CD147 positive color was brown and yellow, and positive position was located mainly in the cytomembrane, and a small amount of cytoplasm was appeared. Among 14 cases of parathyroid carcinoma, 11 cases of CD147 positive score was 3+ , 3 cases of CD147 positive score was 2+ ; Among 33 cases of parathyroid adenoma , 8 cases of CD147 positive score was 2+ , 15 cases of it was 1+ , 10 cases of it was negative. CD147 was highly expressed in parathyroid carcinoma tissues, and the expression of CD147 was significantly different from the expression of parathyroid adenoma(P<0.05). CONCLUSION: CD147 immunohistochemical staining can help to diagnose parathyroid carcinoma.

A simplified intrathymic injection technique for mice.

Intrathymic injection is a common technique used for research concerning immunotolerance induction, gene therapy and T cell development in mice. Traditionally used protocols involve major surgery that exposes the thoracic cavity, which results in injury to the mice and increased risk of poor recovery and postsurgical complications such as infection. We introduce a simplified intrathymic injection technique that does not expose the thoracic cavity and virtually eliminates pain, distress and postoperative complications while maintaining high injection efficiency. The technique is suitable for both adult and neonatal mice.

Evaluation of the cytotoxicity of a two photon absorbing fluorescence compound on human HepG2 cells and its application to tracking human hepatic cancer cells in mice.

Small organic dyes have been applied widely in fluorescence imaging techniques for biomedical research. We investigated the cytotoxicity of a novel fluorescent dye, trans-4-(N-2-hydroxyethyl-N-ethyl amino)-4'-(dimethyl amino) stilbene (DMAHAS), on human hepatocellular carcinoma (HepG2) cells using methyl thiazolyl tetrazolium(MTT), a neutral red assay, a Coomassie brilliant blue assay, and flow cytometric analysis. Our results showed that DMAHAS had live cell permeability, stable cytosolic localization and no significant cytotoxicity to HepG2 cells. We explored its application further for tumor cell tracking in a human liver tumor xenograft mouse model. Tumor xenografts were examined by fluorescence imaging and conventional histological methods. In addition, a method based on DMAHAS release was developed for tumor-specific cytotoxicity analysis. Our study indicated that DMAHAS is a reliable probe for tumor tracking and fluorescence imaging.

Molecular cloning and characterization of cotton cDNAs expressed in developing fiber cells.

Using the FDD-PCR technique, ten new fiber-specific cDNAs were isolated from developing cotton fiber cells and showed high amino acid identity to previously recorded cDNAs. Five cDNAs encoding bisphosphate nucleotidase, alpha-tubulin, beta-galactosidase, annexin, and reversibly glycosylated polypeptide were identified while the functions of five other cDNAs were undetermined. Dot blot analysis showed that all transcripts of the 10 cDNAs accumulated preferentially in fiber cells and the majority were expressed in the early phase of cotton fiber development, except for F14 which accumulated at a high level during the late phase of developing fiber cells, indicating that all of the genes were involved in the process of fiber development.

Components of the ether-insoluble resin glycoside fraction from the seed of Cuscuta australis.

An ether-insoluble resin glycoside fraction was obtained from the seeds of Cuscuta australis. Identification and characterization of alkaline hydrolysis products by means of GC, FABMS, and 1H, 13C, and 2D NMR revealed the material to be composed of three new glycosidic acids, cuscutic acids A1-A3 (1-3); triglycosides with (11S)-jalapinolic or (11S)-convolvulinolic acid as the aglycon; and acetic, isobutyric, (2S)-2-methylbutyric, tiglic, and (2R,3R)-nilic acids. The resin glycoside is considered to be a complex mixture of glycosidic ester-type oligomers (up to heptamers) with a core consisting of a number of the above cuscutic acids each acylated with one or two carboxylic acid moieties.

Components of the ether-insoluble resin glycoside-like fraction from Cuscuta chinensis.

A trisaccharide and four new glycosidic acids, named cuscutic acids A-D, along with known organic acids, acetic acid, propionic acid, (2S)-2-methylbutyric acid, tiglic acid, (2R, 3 R)-nilic acid, (11S) convolvulinolic acid and (11S)-jalapinolic acid have been isolated from the alkaline hydrolysate of the ether-insoluble resin glycoside-like fraction of the seeds of Cuscuta chinensis. The compounds were characterized on the basis of chemical and physical data.