The application of transbronchial lung cryobiopsy guided by endobronchial ultrasound sheath in diagnosis of nonresolving pneumonias / 中国医师进修杂志

Objective:To explore the application of transbronchial lung cryobiopsy guided by endobronchial ultrasound sheath (EBUS-GS-TBCB) in diagnosis of nonresolving pneumonias.Methods:Sixty patients with nonresolving pneumonias from March 2019 to July 2020 in Dalian Municipal Central Hospital were selected. The patients were divided into EBUS-GS-TBCB group (31 cases) and transbronchial forcep lung biopsy guided by endobronchial ultrasound sheath(EBUS-GS-TBLB) group (29 cases) by random digits table method.Results:The diagnostic rate of nonresolving pneumonias in EBUS-GS-TBCB group was significantly higher than that in EBUS-GS-TBLB group: 87.10% (27/31) vs. 65.52% (19/29), and there was statistical difference ( χ2 = 3.90, P = 0.048). There were no statistical difference in sensitivity, specificity, accuracy, positive predictive value and negative predictive value between 2 groups ( P>0.05). There were no statistical difference inthe shortest distance from lesions to pleura, incidence of pneumothorax and incidence of bleeding between EBUS-GS-TBCB group and EBUS-GS-TBLB group: (27.42 ± 2.88) mm vs. (27.01 ± 2.37) mm, 6.45%(2/31) vs. 3.45%(1/29) and 22.58%(7/31) vs. 13.79% (4/29), P>0.05. Among the causes of nonresolving pneumonias, infectious factors accounted for 21.67% (13/60), non infectious factors accounted for 66.67% (40/60), and uncertain causes accounted for 11.67% (7/60). Conclusions:The diagnostic rate of EBUS-GS-TBCB in nonresolving pneumonias is significantly higher than EBUS-GS-TBLB, and the complications such as bleeding and pneumothorax do not increase significantly.

Construction and function of a root-specific promoter SRSP / 生物工程学报

The responsibility of root is absorbing water and nutrients, it is an important plant tissue, but easily to be affected by biotic and abiotic stresses, affecting crop growth and yield. The design of a synthetic root-specific promoter provides candidate promoters for the functional analysis and efficient expression of stress-related genes in crop roots. In this study, a synthetic root-specific module (pro-SRS) was designed using tandem four-copies of root specific cis-acting elements (OSE1ROOTNODULE, OSE2ROOTNODULE, SP8BFIBSP8AIB, and ROOTMOTIFAPOX1), and fused with minimal promoter from the CaMV 35S promoter to synthesize an artificially synthetic SRSP promoter. The SRSP promoter was cloned in pCAMBIA2300.1 by replacing CaMV 35S promoter so as to drive GUS expression. The constructs with SRSP promoter were transformed in tobacco by Agrobacterium-mediated method. SRSP promoter conferred root-specific expression in transgenic tobacco plants through Real-time PCR (RT-PCR) analysis and GUS histochemical staining analysis. It is indicated that the repeated arrangement of cis-acting elements can realize the expected function of the promoter. This study laid a theoretical foundation for the rational design of tissue-specific promoters.

Preparation and purification of Cry1Ah protein candidate reference material / 生物工程学报

With the rapid development of transgenic technology, the safety of genetically modified products has received extensive attention. Certified reference materials for the detection of genetically modified organisms play important roles in ensuring comparability and traceability of the qualitative and quantitative detection of genetically modified products. However, the development of protein reference materials is relatively slow, and one of the difficulties is the preparation of protein candidates with high purity. The cry1Ah1 gene of Bacillus thuringiensis has been used for the development of transgenic insect-resistant crops because of its excellent insecticidal activity against lepidopteran pests such as Asian corn borer, and has obtained transgenic lines with good insect resistance traits. In order to develop Cry1Ah protein certified reference material, it is urgent to establish a preparation and purification system. In this study, a system for preparing Cry1Ah protein by Bt expression system was optimized, and a high-purity Cry1Ah protein (size exclusion chromatography purity: 99.6%) was obtained by ion-exchange chromatography and size exclusion chromatography stepwise purification. The results of biological activity assay showed that there was no significant difference in the insecticidal activity of purified Cry1Ah protein and protoxin against diamondback moths (Plutella xylostella). Finally, the amino acid sequence of the activated Cry1Ah protein was determined using Edman degradation and mass spectrometry. In summary, the obtained Cry1Ah pure protein can be used for the development of protein reference materials.

Clinical and pathological analysis of lobular capillary hemangioma / 中国医师进修杂志

Objective To explore the chnical manifestation,pathological features and characteristics of lobular capillary hemangioma (LCH),so as to raise awareness.Methods Sixty-nine cases of LCH were studied by routine histopathologic and immunohistochemical examinations.Results The age range of the 69 patients including 34 males and 35 females with LCH was 4-80 years,with an average age of 42 years.The lesions of 50 cases were located in head and neck region,and the other 19 cases were in other parts of the body,with 1 case in the left main bronchus and intracalvarium.The average diameter of the tumors was about 1 cm,and parts of the tumors showed polypoid masses.Most of the tumors were in the skin and mucosa.Granulomatous lesions showed the proliferation of thin-walled and thick-walled capillaries in a lobular pattern.The vascular interstitium was edematous and was infiltrated by the inflammatory cells.Immunohistochemical examination showed CD34 positive in the endothelial cells and smooth muscle actin positive in the peri-vascular spindle cells.Conclusions Studies of LCH show the obvious features on age,gender and diseased location,and very few have a relapse.Its pathogenesis is still unclear.LCH is rarely found in the trachea,and is more rarely in the central nervous system.So far,there is no case report to show LCH occurs both in the bronchus and intracalvarium.In the special case,the early lesion is found in the left main bronchus.Although the patient is treated,LCH still recurs for many times in a short period.Finally the tumor is also found in the intracalvarium.Therefore,it's critical to alert whether LCH also occurs in the important organs at the early stage.Complete surgical resection and / or adjuvant radiotherapy can avoid delaying the treatment and the subsequent serious outcomes.

Comparison of codon optimizations of cry1Ah1 gene in rice / 生物工程学报

cry1Ah1, one of holo-type cry genes, cloned in this laboratory from Bacillus thuringiensis strain has been patented in China, and it encoded a protein with strong insecticidal activity against certain lepidopteran insect pests, such as Chilo suppressalis. cry1Ah1 gene is exhibiting good application prospects. In order to improve the expression level of cry1Ah1 gene in rice, and investigate the effect of codon usage preference of gene expression, we designed five different optimized schemes for cry1Ah1 insecticidal critical fragment in accordance with bias of rice codon, to improve G+C content, removed the shear signal and unstable factors. Optimized cry1Ah1 genes were transformed into Escherichia coli Rosetta (DE3) respectively, and 65 kDa polypeptides was expressed normally in inclusion body separately. All of these expressed polypeptides showed insecticidal activity against 2nd-instar larvae of Plutella xylostella and neonate of Chilo suppressalis. After transformation with modified cry1Ah1 genes into Var nippobare, the transgenic rice seedlings were detected by PCR, the positive rate containing target gene was more than 87%. Afterwards, the results of real-time RT-PCR and ELISA assay indicated that the highest expression level of five modified cry1Ah1 genes was that using the highest frequent codons. Average expression amount of Cry1Ah1 polypeptides was 0.104% of total soluble proteins from the positive transgenic rice.

Non-invasive analysis coronary artery flow velocity of slow coronary flow phenomenon / 中华超声影像学杂志

Objective To non-invasive assess coronary blood flow velocity changes of patients with slow coronary flow phenomenon (SCFP) by coronary blood flow imaging (CFI).MethodsTwenty-one patients who had no significant coronary artery stenosis but had thrombolysis in myocardial infarction (TIMI) slow-flow phenomenon were the experimental group,nine patients who has no significant coronary stenosis and TIMI flow normal were the control group.Using corrected TIMI frame count(CTFC) assess velocity of coronary artery.The left ventricular end diastolic diameter,end systolic diameter,ejection fraction,E peak velocity,A peak velocity,E/A ratio were measured by conventional echocardiography.The distal anterior descending coronary artery diastolic peak flow velocity(Vmax),mean velocity(Vmean) and blood flow velocity time integral(VTI) were measured by CFI.Results The corrected TIMI frame count (CTFC) of left anterior descending artery blood flow in slow blood group was (45.37 ± 8.62)frame,that in control group was (15.94± 4.66)frame,the difference was statistically significant (t = -9.596,P =0.000).The conventional echocardiographic measurements of two groups were not significantly different.The left anterior descending artery Vmax was (22.86 ± 3.04)cm/s,Vmean was (17.62 ± 2.89)cm/s,VTIwas (8.49± 2.01)cm in the slow blood flow group,the left anterior descending artery Vmax was (31.78 ± 9.28) cm/s,Vmean was (23.67 ± 7.60) cm/s,VTI was (10.91 ± 4.47) cm in the control group.The difference was statistically significant.The left anterior descending artery CTFC with Vmax and Vmean was negative correlation in the control group and the slow blood flow group.The left anterior descending artery CTFC was negatively correlated with VTI in the control group,there was no correlation between left anterior descending artery CTFC and VTI in the slow blood flow group.Conclusions Coronary artery flow velocity in the left anterior descending artery was declined.CFI can reflect changes in coronary TIMI flow,but in the diagnosis of coronary slow flow phenomenon CFI has limitations.

Effect of clopidogrel premedication on clinical outcomes and bleeding complications in patients undergoing coronary artery bypass graft surgery / 中华急诊医学杂志

Objective To evaluate the effect of clopidogrel premedication on in-hospital major adverse cardiovascular and cerebral events (MACCE) and bleeding outcomes before coronary artery bypass graft surgery (CABG). Method A total of 2021 patients who underwent CABG from July 2003 to September 2005 were divided into either clopidogrel ( n = 479) or no clopidogrel (1542) group before CABG. Patients with clopidogrel administration ( n = 479) were subdivided into < 5 d ( n = 154) ,5～7d(n = 183)and >7d(n = 142) group according to timing of clopidogrel withdrawal before surgery. In-hospital MACCE and perioperative bleeding outcomes were analyzed among groups. Results Patients who took clopidogrel before surgery had nonsignificantly rates of bleeding and in-hospital MACCE compared with those patients not administered clopidogrel. No differences were found about the incidence of total bleeding,minor bleeding,transfusions of red blood cells,fresh frozen plasma,whole blood and in-hosptial MACCE among three subgroups.The < 5 d group had higher incidence of major bleeding and more platelets transfusions than 5 ～ 7 d [47.8% vs. 31.9%,P < 0.017; (0.08 ±0.38) U vs. (0.00±0.00) U,P <0.017,respectively]and >7 d group [47.8% vs. 20.3%,P <0.017; (0.08±0.38) U vs. (0.00±0.00) U,P <0.017,respectively). However,there were no significant differences between 5 ～ 7 d and > 7 d group ( P > 0.05). Conclusions Gopidogrel administration before CABG does not increase the incidence of in-hospital MACCE events. However,the perioperative risk of bleeding will rise if the patients withhold clopidogrel less than five days before surgery.