Analysis of endogenous plasmids in Lacticaseibacillus paracasei ZY-1 and development of expression vectors / 生物工程学报

The construction of efficient and stable Lactobacillus expression vector is critical for strain improvement and development of customized strains. In this study, four endogenous plasmids were isolated from Lacticaseibacillus paracasei ZY-1 and subjected to functional analysis. The Escherichia coli-Lactobacillus shuttle vectors pLPZ3N and pLPZ4N were constructed by combining the replicon rep from pLPZ3 or pLPZ4, the chloramphenicol acetyltransferase gene cat from pNZ5319 and the replicon ori from pUC19. Moreover, the expression vectors pLPZ3E and pLPZ4E with the promoter Pldh3 of lactic acid dehydrogenase and the mCherry red fluorescent protein as a reporter gene were obtained. The size of pLPZ3 and pLPZ4 were 6 289 bp and 5 087 bp, respectively, and its GC content, 40.94% and 39.51%, were similar. Both shuttle vectors were successfully transformed into Lacticaseibacillus, and the transformation efficiency of pLPZ4N (5.23×102-8.93×102 CFU/μg) was slightly higher than that of pLPZ3N. Furthermore, the mCherry fluorescent protein was successfully expressed after transforming the expression plasmids pLPZ3E and pLPZ4E into L. paracasei S-NB. The β-galactosidase activity of the recombinant strain obtained from the plasmid pLPZ4E-lacG constructed with Pldh3 as promoter was higher than that of the wild-type strain. The construction of shuttle vectors and expression vectors provide novel molecular tools for the genetic engineering of Lacticaseibacillus strains.

Clinical value of marker genes in the diagnosis of salivary gland mammary secretory carcinoma / 中华内分泌外科杂志

Objective:To investigate the clinicopathological features, immunophenotype, diagnosis and differential diagnosis of mammary analogue secretary carcinoma of salivary gland (MASC) .Methods:From Jan. 2018 to Jan. 2021, 28 cases of salivary gland MASC were collected in Ningbo Diagnostic Pathology Center, and 10 cases of acinar cell carcinoma and 10 cases of adenoid cystic carcinoma were selected as controls. Immunohistochemical envision method was used to detect the expression of S-100,, DOG-1, CD117, SOX-10, Mammaglobin and Vimentin, and fluorescence in situ hybridization was used to detect the fusion gene of ETV6-NTRK3.Results:The S-100 protein, SOX-10 and Vimentin of MASC of salivary gland were diffusingly positive (28/28) , Mammaglobin (22/28) and CD117 (19/28) were partially positive, and DOG-1 was negative. ETV6-NTRK3 fusion transcription was successfully detected in 26 of 28 salivary gland MASC cases, of which 23 were positive and 3 were negative.Conclusions:Salivary gland MASC is a low-grade malignant epithelium tumor. Comprehensive detection of the expression levels of S-100 protein, SOX-10, DOG-1, Mammaglobin and CD117 is of great value for the diagnosis and differential diagnosis of MASC. FISH detection of ETV6-NTRK3 gene fusion has important reference value for definite diagnosis.

Design of Biochemistry PBL teaching cases suitable for engineering education / 生物工程学报

Engineering education is an important part of China's higher education. With the deepening of the connotation of new engineering talents training, it is timely to carry out the curriculum reform in all directions and improve the quality of engineering talents. In order to highlight the characteristics of fostering new engineering talents, specialized courses and practical courses are becoming the focus of curriculum teaching reform. However to this end, it is urgent to explore how to strengthen fostering talents with engineering characteristics in the professional foundation courses. Taking the biochemistry course as an example, problem-based learning teaching methods were employed, appropriate teaching cases were selected, scientific and technical problems based instructional designs were explored, and students were guided to condense, analyze and solve problems. This study aims to transform the learning attitude of students from "passive" to "active", enhance students' thinking skills, highlight the engineering application of science and technology, lay the foundation for continuous and even lifelong learning, and provide a reference for fostering talents under the new engineering background.

IgG4-related kidney disease presenting as a cystic renal mass: a case report and literature review / 中华泌尿外科杂志

Objective:To investigate the clinical characteristics, differential diagnosis and treatment of IgG4-related kidney disease (IgG4-RKD) presenting as a cystic renal mass.Methods:A 42-year-old male patient was found having a tumor in the left kidney in an annual physical examination. Subsequent CTU scan revealed a round shadow in the lower pole of the left kidney, with slightly lower, uneven density and obscure boundaries, which was slightly enhanced on contrast-enhanced CT imaging, suggesting a cystic mass (Bosniak category Ⅲ). Contrast-enhanced MRI showed a long T1, long T2 signal lesion in the lower pole of the left kidney, which was not obviously enhanced in the cortical phase but exhibited continuous and separated enhancement in parenchymal and delayed phases, suggesting a cystic renal cell carcinoma. A preoperative diagnosis of a left renal mass was made, for which the patient underwent a laparoscopic partial nephrectomy.Results:The resected cystic mass was grey white with obscured boundaries. Microscopically, the mass showed lymphoplasmacytic hyperplasia with lymphoid follicles, cystic formation, fibrosis hyperplasia and vascular obstruction. Immunohistochemistry revealed that the number of IgG4 (+ ) plasma cells was more than 50/HPF, and over 40% IgG (+ ) plasma cells were IgG4 (+ ). By virtue of the radiologic and pathologic findings, the patient was diagnosed with IgG4-RKD. At 2 weeks after operation, serum IgG level was 0.71g/L (reference: 0.03-2.01 g/L). Chest and abdominal CT scans performed 3 months after operation showed no evidence of other organ involvement and serum IgG4 level was within the normal range. Therefore no adjuvant therapy was prescribed.Conclusions:It is usually difficult to differentiate IgG4-RKD presenting as a cystic renal mass by radiography alone. The presence of other organ involvement, serum IgG4 test and renal biopsy can help the diagnosis and so as to avoid unnecessary surgery.

Co-word clustering analysis for hot research areas of human caring based on PubMed / 中华护理杂志

Objective To identify hot research areas of nursing human caring in PubMed from 2012 to 2016, and to explore the present research status and development directions. Methods PubMed was searched using key words human caring. BICOMB 2.0 and SPSS 11.5 software were used to analyze high-frequency keywords and conduct co-word clustering analysis. Results We searched for 3088 related articles and extracted 42 high-frequency keywords (30.97%). Seven hot research areas were identified,including:human caring in nursing practice;nursing models,nursing theories;nursing education of human caring;hospice care;relationship between human caring and nurse-patient relation;human caring of cancer patients;family system,social support and human caring. Conclusion Analysis of research areas of nursing human caring in past 5 years is beneficial to understanding the present research status and development directions,and providing references for practice,research and education of human caring.

Clinicopathological features of multilocular cystic renal cell carcinoma:a series of 32 cases / 临床与实验病理学杂志

Purpose To investigate the clinicopathological features of multilocular cystic renal cell carcinoma ( MCRCC) , and to im-prove the understanding of this disease. Methods Thirty-two of MCRCC were studied by clinic data, pathological features and immu-nophenotype. All the thirty-two cases were followed up. Results In this study, thirty-two patients were diagnosed as MCRCC with a male-to-female ratio of 2.2 : 1, thirteen of whom had the tumor in the left kidney, eighteen in the right kidney, another in the double kidneys. The mean of tumor diameter was 4.6 cm (1.0~8.0 cm). Eleven patients underwent radical nephrectomy while the other 21 patients received nephron sparing surgery. Microscopically, all the cases were multiocular, lined in the cyst wall by a single layer of tumor cells with the clear or pale cytoplasm and Fuhrman grade 1 nuclei. Occasionally, the lining consisted of several layers of tumor cells or a few small papillae were present. The linings of the cyst wall were rich in thin-wall blood vessels. Immunohistochemically, the tumor cells were positive for CK(32/32), CK7(25/32), EMA(32/32), CD10(23/32) and vimentin(20/32), while negative for CD68. Conclusions MCRCC is characteristic by low grade nuclei, lacking solid nodules, and in the cyst wall, and has a favourable prognosis. The rich in thin-wall blood vessels and the lining cells with the clear or pale cytoplasm are diagnosis clues.

Improving Potato Plants Oxidative Stress and Salt Tolerance by Gene Transfer Both of Cu/Zn Superoxide Dismutase and Ascorbate Peroxidase / 中国生物工程杂志

In plants, oxidative stress is one of the major causes of damage as a result of various environmental stresses and it is primarily due to the excessive accumulation of reactive oxygen species. To develop transgenic potato plants with enhanced tolerance to environmental stress, transgenic potato plants (Solanum tuberosum L. cv. Atlantic) expressing the Cu/ZnSOD and APX genes in chloroplasts were generated under the control of the oxidative stress-inducible promoter. To investigate oxidative stress tolerance, transgenic plants were evaluated at the level of leaf discs and plantlets after methyl viologen (MV) and salt treatment. Leaf discs from transgenic potato plants showed 13% less membrane damage compared to non-transgenic (NT) plants suffering 10 μmol/L MV treatment of 48 h, and showed 1.6-fold higher chlorophyll contents than those of NT plants at 1.0mol/L NaCl treatment (31% vs. 19%). In addition, transgenic potato plants maintained higher rooting rates (75%) during 100mmol/L NaCl treatment than those (12%) from NT plants. Moreover, the tolerance to salt stress in transgenic plants was consistent to increased transcript levels and higher activities of SOD and APX compared to NT plants. These results suggest that expression of Cu/ZnSOD and APX in chloroplasts could be used in plants to enhance the tolerance to environmental stresses.

Agrobacterium-mediated Transformation of an Economically Important Potato Cultivar Using Internodal Stem Explants / 中国生物工程杂志

Potato cultivar Atlantic is widely grown for potato chips in the world. However, this economically important potato cultivar exhibits very poor yields and traits under severe environmental stress. To develop an efficient plant transformation system that could be used to produce large scale transgenic potato plants with enhanced tolerance to environmental stress and therefore would be beneficial for potato processing industry, Agrobacterium-mediated transformation of internodal stem explants using both superoxide dismutase (SOD) and ascorbate peroxidase (APX) genes under the control of an oxidative stress-inducible SWPA2 promoter was performed. Comparing to leaf explants, stem internodal explants were less liable to damage during manipulation, more amenable to in vitro conditions. The addition of silver thiosulfate to the selection medium considerably promoted the shoot induction from explant-derived callus. Seven to nine shoots per stem explant were obtained. By combining the best treatments, this system yielded shoot induction frequency of 94.2% and transformation frequency of 80% of internodal stem explants. Stable integration of the transgenes was confirmed by PCR and Southern blot analyses. In conclusion, short duration (7～8 weeks), high efficiency and easy process make this system well suited for wider commercial applications of transgenic Atlantic potato plants.