The electrical impedance spectroscopy (EIS) was analyzed to one-year-old needles of Pinus bungeana Zucc.in an 8-year provenance field trial at the Nursery in Beijing Thirteen Tombs with provenances of Mangshan of Beijing, Xiaoyi of Shanxi, and Liangdang of Gansu. The parameters of EIS and the activities of superoxide dismutase (SOD), the activities of peroxidase (POD), the content of malondialdehyde (MDA), the content of soluble protein, the content of proline, the chlorophyll content and the content of dry matter in non-frost exposed needles were tested with different provenances. The relations between EIS and physiological parameters were analyzed. The frost hardiness (FH) of needles was measured by means of the visual scoring of damage (VSD) after artificial freezing test. The results showed that: (1) Both the EIS parameters and the physiological parameters changed during frost hardening, and the EIS parameters and some physiological parameters differed significantly in certain stages of frost hardening, especially in the early stage, which was in accordance with the differences of FH in different provenances. (2) The EIS parameters membrane time constant τm and extracellular resistance re correlated negatively with the chlorophyll content (the correlation coefficient r were -0.91 and -0.68, respectively), the β correlated negatively with the dry matter content (r=-0.74), while the intracellular resistance ri and re had positive correlation with the dry matter content (r were 0.69 and 0.58, respectively), as well as the β with the MDA content (r=0.68). (3) The β correlated positively with the FH assessed by the VSD method (r=0.85), whereas the ri and re did negatively (r were -0.79 and -0.66, respectively).
To clone and express the malate dehydrogenase (MDH) gene of Toxoplasma gondii, and analyze the immunogenicity.Total RNA was extracted from tachyzoites of RH strain of T. gondii (GenBank accession No. AY650028). The coding region of TgMDH was amplified with a pair of specific primers. The product of RT-PCR was digested with double restriction enzyme and ligated into pET30a (+) vector. The recombinant pET30a (+)-TgMDH plasmid was transformed into E. coli DH5alpha. The positive clones were confirmed by the double restriction enzyme digestion, PCR and sequencing. The correct plasmid was transformed into E. coli BL21 and induced by IPTG. The expressed proteins were analyzed by SDS-PAGE. Conditions for expression were optimized. Abundant soluble rTgMDH protein was purified with Ni-NTA affinity chromatography. Mice was intranasally immunized with purified rTgMDH and murine anti-rTgMDH serum was prepared. Western blotting with murine anti-rTgMDH serum and rabbit anti-T. gondii serum was used to analyze its immunogenicity.The product of RT-PCR was with 951 bp. The recombinant plasmid pET30a(+)-TgMDH was confirmed by the double restriction enzyme digestion, PCR and sequencing. A soluble recombinant protein with relative molecular weight of 36 000 was analyzed by SDS-PAGE, followed by coomassie blue staining. Western blotting revealed that rTgMDH can be recognized by murine anti-rTgMDH serum and rabbit anti-T. gondii serum.TgMDH gene has been expressed in prokaryotic expression system and shows immunogenicity.
// Dandan Yu 1, 9, 10 , Yali Zhong 1 , Xiaoran Li 9, 10 , Yaqing Li 1 , Xiaoli Li 1 , Jing Cao 1, 2 , Zhirui Fan 1 , Huijie Fan 1 , Long Yuan 3 , Benling Xu 4 , Yuan Yuan 5 , Hongquan Zhang 6 , Zhenyu Ji 7 , Jian-Guo Wen 8 , Mingzhi Zhang 1 , Jahn M. Nesland 9, 10 , Zhenhe Suo 1, 9, 10 1 Department of Oncology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China 2 Departments of Pathology, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China 3 Department of Surgery, The Affiliated Cancer Hospital of Zhengzhou University, Zhengzhou, Henan, China 4 Department of Cancer Biotherapy, The Affiliated Cancer Hospital of Zhengzhou University, Zhengzhou, Henan, China 5 Department of Pathology, Capital Medical University, Beijing, Fengtai, China 6 Department of Anatomy, Histology and Embryology, Peking University Health Science Center, Beijing, Fengtai, China 7 Department of Oncology, Henan Academy of Medical and Pharmaceutical Sciences, Zhengzhou University, Zhengzhou, Henan, China 8 Institute of Clinical Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou University, Zhengzhou, Henan, China 9 Department of Pathology, The Norwegian Radium Hospital, Oslo University Hospital, Oslo, Norway 10 Department of Pathology, Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Oslo, Norway Correspondence to: Zhenhe Suo, email: zhenhes@medisin.uio.no Keywords: TALEN, fumarate hydratase, gene technology, obesity, heterozygote Received: February 23, 2016 Accepted: August 09, 2016 Published: August 19, 2016 ABSTRACT Transcription activator-like effector nucleases (TALENs) are valuable tools for precise genome engineering of laboratory animals. Here we utilized this technique for efficient site-specific gene modification to create a fumarate hydratase (FH) gene knockout rat model, in which there was an 11 base-pair deletion in the first exon of the FH gene in 111 rats. 18 live-born targeted mutation offsprings were produced from 80 injected zygotes with 22.5% efficiency, indicating high TALEN knockout success in rat zygots. Only heterozygous deletion was observed in the offsprings. Sixteen pairs of heterozygous FH knockout (FH+/–) rats were arranged for mating experiments for six months without any homozygous KO rat identified. Sequencing from the pregnant rats embryo samples showed no homozygous FH KO, indicating that homozygous FH KO is embryonically lethal. Comparatively, the litter size was decreased in both male and female FH+/– KO rats. There was no behaviour difference between the FH+/– KO and the control rats except that the FH+/– KO male rats showed significantly higher body weight in the 16-week observation period. Clinical haematology and biochemical examinations showed hematopoietic and kidney dysfunction in the FH+/– KO rats. Small foci of anaplastic lesions of tubular epithelial cells around glomeruli were identified in the FH+/– kidney, and these anaplastic cells were comparatively positive for Ki67, p53 and Sox9, and such findings are most probably related to the kidney dysfunction reflected by the biochemical examinations of the rats. In conclusion, we have successfully established an FH+/– KO rat model, which will be useful for further functional FH studies.
The incidence of synchronous multiple primary malignancies has been reported to be low. We report a rare case of synchronous lung squamous cell cancer and small cell lung cancer in an 82-year-old male patient. There is a lack of standard diagnostic criteria for multiple primary lung cancer. Two tumors with similar morphology are difficult to draw conclusions about the same lineage or different lineages. If the patient's physical condition permits, multiple tumors should be sampled and tested. Besides, imaging features are helpful for identification. It is advisable to diagnose synchronous multiple primary malignancies in an early stage, which contributes to a favorable outcome.
To observe the reproductive modes of Blastocystis hominis and study the relation between this protozoa and bacteria.Using the Iodine and Haematoxylin staining, the morphology of B. h from patients and RPMI 1640 medium were observed. The B. h positive mucous diarrheal specimens were cultured and identified any possible known pathogenic intestinal bacteria. B. h and colibacillus were co-cultured to observe the interaction between them.Four modes of reproduction for B. h were confirmed: binary fission, endodyogeny, multiple fission and budding. The fact that there was no other intestinal pathogens in half of the B. h positive specimens suggested B. h may cause disease independently. B. h and colibacillus were restrained each other.B. h reproduces in at least four modes. B. h could be a pathogen and its pathogenesis may be related to micro-ecological changes.
Increased expression of galectin-1 (Gal-1) in carcinoma-associated fibroblasts (CAFs) has been reported to correlate with progression and prognosis in many cancers. However, rarely have reports sought to determine whether high Gal-1 expression in CAFs in gastric cancer is involved in the tumor process, and the specific mechanism by which it promotes the evolution of gastric cancer is still unknown. In this study, we cultured gastric cancer CAFs, which showed strong expression of Gal-1, and established a co-culture system of CAFs with gastric cancer cells. Specific siRNA and in vitro migration and invasion assays were used to explore the effects of the interaction between Gal-1 expression of CAFs and gastric cancer cells on cell migration and invasion. We found that the overexpression of Gal-1 in CAFs enhanced gastric cancer cell migration and invasion, and these stimulatory effects could be blocked by specific siRNA which reduced the Gal-1 expression level. A set of cancer invasion-associated genes were then chosen to identify the possible mechanism of Gal-1-induced cell invasion. Among these genes, integrin β1 expression in cancer cells was considered to be associated with Gal-1 expression. Pre-blocking of the integrin β1 expression in gastric cancer cells with siRNA could interrupt the invasion-promoting effect of CAFs with high Gal-1 expression. Furthermore, immunohistochemical assay confirmed a positive correlation between Gal-1 and integrin β1 expression. Our results showed that high expression of Gal-1 in CAFs might facilitate gastric cancer cell migration and invasion by upregulating integrin β1 expression in gastric cancer.
'/%3e%3cuse xlink:href='%23a' transform='rotate(23.036 .093 25.536)'/%3e%3cuse xlink:href='%23a' transform='rotate(45.87 1.273 16.18)'/%3e%3cuse xlink:href='%23a' transform='rotate(69.945 .996 12.078)'/%3e%3cuse xlink:href='%23a' transform='rotate(20.66 -19.689 31.932)'/%3e%3c/svg%3e)