Investigation on the design, production and application of human anatomy micro-course in the information age / 中华医学教育探索杂志

Taking the micro-course The Connection Between Vertebrae as an example, this paper discusses the teaching design, production process and practical application of the anatomy micro-course, analyzes the existing problems and puts forward corresponding suggestions. The micro-course design integrates knowledge points with ideological and political elements, and applies a variety of teaching methods to make the teaching content interesting, enlightening and applicable. In the production process, using anatomical specimens, models and 3D software demonstration structure can make abstract knowledge intuitive and perceptual. Moreover, the artistry of micro-course is increased appropriately, and the misunderstanding of emphasizing appearance and neglecting design is avoided. The application of micro-course integrates the micro-course and online education platform to realize the intelligent education of information teaching. In addition, college teachers should improve the awareness of micro-course production and carry out systematic construction and application of micro-course resources.

Acupuncture for delayed sleep-wake phase disorder: a randomized controlled trial / 中国针灸

OBJECTIVE@#To observe the clinical effect of acupuncture for delayed sleep-wake phase disorder (DSWPD).@*METHODS@#A total of 84 patients with DSWPD were randomized into an observation group (42 cases, 2 cases dropped off) and a control group (42 cases, 3 cases dropped off). On the basis of sleep hygiene education, acupuncture was applied at Shenmai (BL 62), Zhaohai (KI 6), Hegu (LI 4), Taichong (LR 3), Zusanli (ST 36) and Sanyinjiao (SP 6) in the observation group, while placebo acupuncture was applied at the same acupoints in the control group. The treatment lasted for 8 weeks, once every other day, 3 times a week in the 1st to 4th weeks; once every 3 days, 2 times a week in the 5th to 8th weeks. Before and after treatment, the actigraphy (ACT) indexes of objective sleep (total time of stay in bed, total sleep time, sleep efficiency, the number of awakenings and the wake time after falling asleep) and plasma cortisol (CORT) level were observed; before and after treatment and in follow-up of 1, 3 months after treatment, the scores of morningness-eveningness questionnaire (MEQ), insomnia severity index (ISI), fatigue severity scale (FSS) and Epworth sleepiness scale (ESS) were observed in the two groups.@*RESULTS@#Compared before treatment, the total sleep time was prolonged, the sleep efficiency was improved, the number of awakenings was reduced, and the wake time after falling asleep was shortened after treatment in the observation group (P<0.01, P<0.05), and those in the observation group after treatment were superior to the control group (P<0.01, P<0.05). Compared before treatment, the MEQ scores after treatment in both groups and in the follow-up of 1, 3 months after treatment in the observation group were increased (P<0.01), and the MEQ score of each time point after treatment in the observation group was higher than the control group (P<0.01). The scores of ISI, FSS and ESS after treatment, and the scores of ISI、ESS in follow-up of 1, 3 months after treatment in the observation group were decreased compared with those before treatment (P<0.01, P<0.05), and in the observation group, the scores of ISI, FSS and ESS of each time point after treatment were lower than those in the control group (P<0.01, P<0.05). After treatment, the plasma CORT level in the observation group was decreased compared with that before treatment and that in the control group (P<0.01, P<0.05).@*CONCLUSION@#Acupuncture can improve the sleep and wake phase of patients with DSWPD, improve sleep quality and daytime function, and its mechanism may be related to the down-regulation of plasma CORT level.

Clinicopathological Features of Primary Small Cell Neuroendocrine Carcinoma of the Bladder / 中国医学科学院学报

Objective To investigate the clinicopathological features,immunohistochemical features,diagnosis,and relationship with sporadic prostate cancer in primary small cell neuroendocrine carcinoma of the bladder. Methods We retrospectively analyzed the clinical characteristics of 12 patients with primary small cell neuroendocrine carcinoma of the bladder diagnosed at Beijing Chao-Yang Hospital affiliated to Capital Medical University from January 2013 to September 2022.The histological features of primary small cell neuroendocrine carcinoma of the bladder were re-evaluated by two pathologists according to the 2022 revision of the World Health Organization Classification of Tumors of the Urinary System and Male Genital Organs.Electronic medical records were retrieved,and telephone follow-up was conducted from the time of histopathological diagnosis to the death or the end of the last follow-up until January 31,2023. Results The 12 patients include 7 patients in pT3 stage and 1 patient in pT4 stage.Eight patients were complicated with other types of tumors,such as high-grade urothelial carcinoma of the bladder and squamous cell carcinoma.Five patients had sporadic prostate cancer.Immunohistochemical staining showed that 12 (100.0%),10 (83.3%),and 8 (66.7%) patients were tested positive for CD56,Syn,and CgA,respectively.The Ki67 proliferation index ranged from 80% to 90%.Five patients with urothelial carcinoma were tested positive for CK20,GATA3,and CK7.P504S was positive in all the 5 patients with prostate cancer,while P63 and 34βE12 were negative.The follow-up of the 12 patients lasted for 3-60 months.Eight of these patients died during follow-up,with the median survival of 15.5 months.Four patients survived. Conclusions Primary small cell neuroendocrine carcinoma of the bladder is a rare urological tumor with high aggressiveness and poor prognosis.In male patients with bladder prostatectomy,all prostate tissue should be sampled.If prostate cancer is detected,the prostate-specific antigen level should be monitored.

Analysis of chemical constituents in ethyl acetate extract of Taxilli Herba by UPLC-Q-Exactive-MS and screening of potential xanthine oxidase inhibitors / 中国中药杂志

The present study analyzed and identified the chemical constituents from ethyl acetate(EA) extract of Taxilli Herba with UPLC-Q-Exactive-MS and screened active xanthine oxidase(XO) inhibitors with HPLC. The analysis was performed on an Hypersil GOLD C_(18) reversed-phase column(2.1 mm×50 mm, 1.9 μm), with the mobile phase of water containing 1% formic acid(A) and methanol(B) under gradient elution, the flow rate of 0.3 mL·min~(-1), and the injection volume of 5 μL. ESI source was used for MS and the compounds were collected in positive and negative ion modes. Xcalibur 4.1 was used to analyze the retention time, accurate relative molecular weight, and fragmentation of the compounds. The inhibitory activity of some known compounds on XO was screened by HPLC. Thirty chemical constituents were identified, including phenolic acids and flavonoids by experimental data combined with information of standards, data reported previously, and databases, such as MzCloud and ChemSpider. The activities of 10 chemical components were screened. Gallic acid and naringenin chalcone had strong inhibitory activities on XO with IC_(50) of 57 μg·mL~(-1) and 108 μg·mL~(-1). UPLC-Q-Exactive-MS allows the accurate, rapid, and comprehensive identification of main chemical constituents from Taxilli Herba. Gallic acid and naringenin chalcone may be the active components of XO inhibitors.

Practice and thinking of online teaching in human anatomy based on "Zhihuishu and Tencent classroom" / 中华医学教育探索杂志

During the outbreak of coronavirus disease 2019 (COVID-19), combining clinical "5+3" integrated students' characteristics and the contents of the courses, teachers applied the Zhihuishu platform and Tencent classroom in anatomy theory teaching to construct an efficient, accurate and interactive online flipped intelligent classroom covering before class -in class -after class. The ARCS (attention, relevance, confidence, satisfaction) motivation model was introduced into the teaching design to help ensure the effective implementation of flipped classroom. Before class, teachers used the Zhihuishu platform to publish learning resources which students can learn independently. In class, teachers used Tencent classroom for live teaching, to explain key and difficult points, and answer and discuss questions. After class, teachers used the Zhihuishu platform to publish exercise tests and assignments. The practice shows that the online teaching mode has been widely recognized by the students. It's helpful to cultivate the learning interest, improve the ability of autonomous learning and the ability of analyzing and solving problems. It also contributes to the implementation of the diversified formative assessment, and it lays a foundation for developing the mixed teaching after the epidemic.

Establishment and evaluation of a cadaveric model of chronic strain-induced upper cervical spine instability based on fascia-bone theory / 中国组织工程研究

BACKGROUND: Studies regarding the instability of upper cervical spine in cadaveric specimens are mostly based on an acute violence model, and there is still no chronic strain-induced upper cervical instability model. OBJECTIVE: To construct and evaluate the cadaveric model of the upper cervical spine instability caused by chronic strain according to the theory of “fascia and bone.” METHODS: Nine fresh cadaveric cervical spine specimens were prepared and the normal vertebral range of motion was detected by Motion Analysis Motion Capture System. The upper cervical spine instability model was constructed by using BOSE dynamic fatigue testing machine with maximum flexion, extension and rotation fatigue loading. Then, the vertebral range of motion was tested again. The anterior flexion, posterior extension, left flexion, right flexion, left and right rotation of the occipitoatlantoaxial joint, the atlantoaxial joint and the whole upper cervical spine were compared before and after modeling. The implementation of the study protocol complied with the relevant ethical requirements of Southern Medical University, and the specimens were voluntarily donated. RESULTS AND CONCLUSION: (1) During anterior flexion, the range of motion of the atlantoaxial joint (C1-2) and the entire upper cervical vertebra (C0-2) of the specimens after modeling was significantly larger than that before modeling (P 0.05). (3) During right rotation, the range of motion of the whole upper cervical spine (C0-2) of the specimens after modeling was significantly increased compared with that before modeling (P < 0.05). During both left and right rotation, the range of motion of the atlantoaxial joint (C1-2) and the whole upper cervical spine (C0-2) of the specimens was significantly larger than that of the specimens before modeling (P < 0.05). Therefore, this model can be used to reflect the state of upper cervical instability caused by chronic strain.

Practice and thinking of online teaching in human anatomy based on "Zhihuishu and Tencent classroom" / 中华医学教育探索杂志

During the outbreak of coronavirus disease 2019 (COVID-19), combining clinical "5+3" integrated students' characteristics and the contents of the courses, teachers applied the Zhihuishu platform and Tencent classroom in anatomy theory teaching to construct an efficient, accurate and interactive online flipped intelligent classroom covering the before class, in class and after class. The ARCS motivation model was introduced into the teaching design to help ensure the effective implementation of flipped classroom. Before class, teachers used the Zhihuishu platform to publish learning resources which students can learn independently. In class, teachers used Tencent classroom for live teaching, to explain key and difficult points, and answer and discuss questions. After class, teachers used the Zhihuishu platform to publish exercise tests and assignments. The practice shows that the online teaching mode has been widely recognized by the students. It’s helpful to cultivate the learning interest, improve the ability of autonomous learning and the ability of analyzing and solving problems. It also contributes to the implementation of the diversified formative assessment, and it lays a foundation for developing the mixed teaching after the epidemic.

Low Doses of Nonylphenol Promote Growth of Colon Cancer Cells through Activation of ERK1/2 via G Protein‒Coupled Receptor 30 / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Nonylphenol (NP) is an endocrine disruptor found in products such as cleaners, plastics, and detergents. It exerts actions similar to endogenous 17β-estradiol (E2) and is reported to influence various cancers. However, its role in colon cancer remains elusive. MATERIALS AND METHODS: Colon cancer cell lines COLO 205 and SW480 were employed in our study. The cells were treated with NP or E2 followed by measurement of apoptosis and proliferation using flow cytometry and MTT assays, respectively. G protein–coupled estrogen receptor 30 (GPR30) expression was visualized using immunofluorescence and Western blot. To investigate the underlying mechanism, the expression levels of GPR30, p-protein kinase A (PKA), c-myc, cyclin D1, and ERK1/2 were analyzed using Western blot. Meanwhile, the GPR30 antagonist G15 was utilized to validate the role of GPR30 in colon cancer progression. Finally, the effect of a GPR30 inhibitor on tumor growth was determined in vivo using tumor xenograft mouse models. RESULTS: NP facilitated the proliferation of colon cancer cells and induced apoptosis failure in vitro. Western blot revealed increased GPR30 expression levels in response to NP treatment. Cyclin D1, p-PKA, c-myc, and proliferating cell nuclear antigen, proteins that regulate the cell cycle, were all upregulated by NP, and NP-mediated ERK1/2 activation and subsequent cell proliferation were abrogated by the GPR30 inhibitor G15. Moreover, colon cancer mice that received G15 administration demonstrated impaired tumor growth in vivo. CONCLUSION: Low dose NP promotes the growth of colon tumors through GPR30-mediated activation of ERK1/2 signaling.

Simultaneous determination of nine constituents in Jiangtangjia Tablets by HPLC / 中成药

AIM To establish an HPLC method for the simultaneous content determination of catalpol,acteoside,jionoside B1,martinoside,calycosin-7-O-β-D-glucoside,ononin,calycosin,formononetin and heterophyllin B in Jiangtangjia Tablets (Rehmanniae Radix,Astragali Radix,Pseudostellariae Radix,etc.).METHODS The analysis of methanol extract of this drug was performed on a 30 ℃ thermostatic Agilent TC-C18 column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1％ glacial acetic acid flowing at 0.9 mL/min in a gradient elution manner,and the detection wavelengths were set at 203,210,254,330 nm.RESULTS Nine constituents showed good linear relationships within their own ranges (r ＞ 0.999 0),whose average recoveries were 96.86％-100.30％ with the RSDs of 0.74％-1.68％.CONCLUSION This stable and reliable method can be used for the quality control of Jiangtangjia Tablets.

Interactions of highly pathogenic avian influenza H5N1 nucleoprotein with NF-κB-inducing kinase(NIK) / 军事医学

Objective To study the interactions between the highly pathogenic avian influenza H5N1 nucleoprotein (H5N1 NP) and NF-κB-inducing kinase (NIK),and to reveal the effect of H5N1 NP on NIK-induced NF-κB transcriptional activity.Methods The gene encoding NIK protein was amplified by RT-PCR from total RNA of HeLa cell line.Eukaryotic expression plasmid pCMV-Myc-NIK and prokaryotic expression plasmid pGEX-4T-1-NP (GST-NP) were constructed by cloning from HeLa cell cDNA and pcDNA3-Flag-NP vector,respectively.Co-immunoprecipitation (co-IP) and GST pull-down were used to test the interactions between H5N1 NP and NIK.Dual-luciferase reporter gene analysis system was used to test the effect of H5N1 NP on NIK-induced NF-κB transcriptional activity.Results Co-IP and GST pull-down showed that pCMV-Myc-NIK and pGEX-4T-1-NP (GST-NP) could express Myc tagged NIK protein and GST tagged NP protein in HEK293T cells and E.coli,respectively,and that H5N1 NP was associated with NIK in vivo and in vitro.Dual-luciferase reporter gene analysis suggested that H5N1 could inhibit NIK-induced NF-κB transcriptional activity.Conclusion H5N1 NP interacts with NIK and inhibits NIK-induced NF-κB transcriptional activity.This finding can facilitate further study of H5N1.

Laparoscopic Versus Open Resection of Small Bowel Gastrointestinal Stromal Tumors: Systematic Review and Meta-Analysis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Laparoscopic resection (LAP) for small bowel gastrointestinal stromal tumors (GISTs) is not as common as for stomach. This study aimed to evaluate the safety and efficacy of LAP for small bowel GISTs with systematic review and meta-analysis.</p><p><b>METHODS</b>The Web of Science, Cochrane Library, Embase, and PubMed databases before December 2016 were comprehensively searched to retrieve comparative trials of LAP and conventional open resection (OPEN) for GISTs of small bowel with a relevance of review object. These researches reported intraoperative and postoperative clinical course (operation time, blood loss, time to first flatus and oral intake, hospital stay, morbidity, and mortality), oncologic outcomes, and long-term survival status.</p><p><b>RESULTS</b>Six studies involving 391 patients were identified. Compared to OPEN, LAP had associated with a shorter operation time (weighted mean difference [WMD] = -27.97 min, 95% confidence interval [CI]: -49.40--6.54, P < 0.01); less intraoperative blood loss (WMD = -0.72 ml; 95% CI: -1.30--0.13, P = 0.02); earlier time to flatus (WMD = -0.83 day; 95% CI: -1.44--0.22, P < 0.01); earlier time to restart oral intake (WMD = -1.95 days; 95% CI: -3.31--0.60, P < 0.01); shorter hospital stay (WMD = -3.00 days; 95% CI: -4.87--1.13, P < 0.01); and a decrease in overall complications (risk ratio = 0.56, 95% CI: 0.33-0.97, P = 0.04). In addition, the tumor recurrence and long-term survival rate showed that there was no significant difference between the two groups of patients.</p><p><b>CONCLUSIONS</b>LAP for small bowel GISTs is a safe and feasible procedure with shorter operation time, less blood loss, less overall complications, and quicker recovery. Besides, tumor recurrence and the long-term survival rate are similar to open approach. Because of the limitations of this study, methodologically high-quality studies are needed for certain appraisal.</p>

Clinical Auditory Phenotypes Associated withGene Mutations in Familial Hypoparathyroidism-deafness-renal Dysplasia Syndrome / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Hypoparathyroidism-deafness-renal dysplasia (HDR) syndrome is an autosomal dominant disorder primarily caused by haploinsufficiency of GATA binding protein 3 (GATA3) gene mutations, and hearing loss is the most frequent phenotypic feature. This study aimed at identifying the causative gene mutation for a three-generation Chinese family with HDR syndrome and analyzing auditory phenotypes in all familial HDR syndrome cases.</p><p><b>METHODS</b>Three affected family members underwent otologic examinations, biochemistry tests, and other clinical evaluations. Targeted genes capture combining next-generation sequencing was performed within the family. Sanger sequencing was used to confirm the causative mutation. The auditory phenotypes of all reported familial HDR syndrome cases analyzed were provided.</p><p><b>RESULTS</b>In Chinese family 7121, a heterozygous nonsense mutation c.826C>T (p.R276*) was identified in GATA3. All the three affected members suffered from sensorineural deafness and hypocalcemia; however, renal dysplasia only appeared in the youngest patient. Furthermore, an overview of thirty HDR syndrome families with corresponding GATA3 mutations revealed that hearing impairment occurred earlier in the younger generation in at least nine familial cases (30%) and two thirds of them were found to carry premature stop mutations.</p><p><b>CONCLUSIONS</b>This study highlights the phenotypic heterogeneity of HDR and points to a possible genetic anticipation in patients with HDR, which needs to be further investigated.</p>

Is Laparoscopic Hepatectomy a Safe, Feasible Procedure in Patients with a Previous Upper Abdominal Surgery? / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Laparoscopic liver resection has become an accepted treatment for liver tumors or intrahepatic bile duct stones, but its application in patients with previous upper abdominal surgery is controversial. The aim of this study was to evaluate the feasibility and safety of laparoscopic hepatectomy in these patients.</p><p><b>METHODS</b>Three hundred and thirty-six patients who underwent laparoscopic hepatectomy at our hospital from March 2012 to June 2015 were enrolled in the retrospective study. They were divided into two groups: Those with previous upper abdominal surgery (PS group, n = 42) and a control group with no previous upper abdominal surgery (NS group, n = 294). Short-term outcomes including operating time, blood loss, hospital stay, morbidity, and mortality were compared among the groups.</p><p><b>RESULTS</b>There was no significant difference in median operative duration between the PS group and the NS group (180 min vs. 160 min, P = 0.869). Median intraoperative blood loss was same between the PS group and the control group (200 ml vs. 200 ml, P = 0.907). The overall complication rate was significantly lower in the NS group than in the PS group (17.0% vs. 31.0%, P = 0.030). Mortality and other short-term outcomes did not differ significantly between groups.</p><p><b>CONCLUSIONS</b>Our study showed no significant difference between the PS group and NS group in term of short-term outcomes. Laparoscopic hepatectomy is a feasible and safe procedure for patients with previous upper abdominal surgery.</p>

Effect of Osthole on Adrenocortical Function in Y1 Mouse Adrenocortical Tumor Cells / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effect of osthole (Ost) on adrenocortical function in Y1 mouse adrenocortical tumor cells.</p><p><b>METHODS</b>Y1 mouse adrenocortical tumor cells were taken as subjects in this experiment. In 10.0%, 1.0%, and 0.1% serum DMEM-F12 medium, Y1 cells were treated with 1, 10, 25, 50, 100, and 200 micromol/L Ost for 24 and 48 h. 0.1% Dimethyl Sulfoxide (DMSO) was taken as negative control group and 1 mmol/L (Bu) 2cAMP as positive control group. Cell growth morphology was observed under inverted microscope. Contents of corticosterone were tested by ELISA. Expression levels of steroids synthase such as Star, Cyp11a1, Cyp21a1, Hsd3b2, Cyp11b1, Cyp11b2, Cyp17a1, and Hsd17b3 mRNA were detected by Real time quantitative PCR (RT-qPCR).</p><p><b>RESULTS</b>Y1 cell proliferation was obviously inhibited by 100 and 200 micromol/L Ost, and its inhibitory effect was more significant in 0.1% serum medium. Compared with the negative control group, gene expressions of Star, Cyp11a1 , Cyp21a1, Hsd3b2, Cyp11b1, Cyp17a1, and Hsd17b3 were significantly enhanced in the posi- tive control group (P < 0.05). Y1 cell corticosterone levels significantly increased in 50 micromol/L Ost treatment group after 24-and 48-h intervention (P < 0.05). Contents of corticosterone increased more obviously in 25 and 50 +/- mol/L Ost treatment groups after 48-h intervention, as compared with 24-h intervention (P < 0.01). After 24-h intervention, expression levels of Star, Cyp21a1, and Hsd3b2 genes were significantly up-regulated in 25 and 50 lLmol/L Ost groups (P < 0.05). Star gene expression was further enhanced after 48-h intervention (P < 0.05). However, Ost showed no effect on Cyp11a1 (P > 0.05). Additionally, gene expressions of Cyp11b1 and Cyp17a1 were significantly enhanced by 10, 25, and 50 pLmolIL Ost after treatment for 24 and 48 h (P < 0.05). Ost showed no obvious effect on Cyp11b2 and Hsd17b3 expressions.</p><p><b>CONCLUSION</b>Ost could regulate adrenal cortex function and promote corticosterone synthesis and secretion through strengthening gene expressions of steroidogenic enzymes.</p>

Construction and preliminary study of biological functions of O157∶H7 typeⅢsecre-tion system effector NleF gene knockout mutant / 军事医学

Objective To construct Escherichia coli O157∶H7 T3SS effector NleF gene knockout mutant and its com-plementary strain, and probe its effects on bacterial growth and cell death .Methods T3SS Effector NleF gene knockout mutant ΔnleF was constructed with λ-Red homologous recombination .Complementary strain ΔnleF/NleF was constructed by transferring pET-24a(+)-NleF into ΔnleF competent cells.Wild type,ΔnleF and ΔnleF/NleF were cultured in LB and DMEM(10%FBS) respectively,D600 was measured every hour , and the growth curve was drawn .HeLa cells were infected with three kinds of strains , the supernatant of LDH release was detected with cytotoxicity detection kit ,and the cytotoxicity was calculated .Results ΔnleF and ΔnleF/NleF were constructed .The growth rates of wild type , ΔnleF and ΔnleF/NleF was not significantly different .Wild type O157 infection induced cell death .Cytotoxicity was increased as much in ΔnleF in-fected cells as in ΔnleF/NleF infected cells.Conclusion EHEC O157∶H7 T3SS Effector NleF has no significant effect on bacterial growth ,but might inhibit host cell death caused by bacterial infection .

Effect of typeⅢsecretion system deficient mutant of E.coli O157∶H7 on attachment of HeLa cells / 军事医学

Objective To construct the typeⅢ secretion system (T3SS) deficient mutant of O157∶H7 EDL933, and to detect its ability of attachment after infection with HeLa cells.Methods The recombinant DNA fragments obtained kana-mycin resistant gene were constructed by overlap extension PCR and transferred into EDL933/pKD46 to replace escR gene by homologous recombination.Results The T3SS-deficient mutant (ΔescR) was successfully construted.Compared with the wild-type EDL933, the growth rate and attachment on HeLa cells of ΔescR were significantly decreased.Conclusion The deletion of escR gene, which encodes T3SS structural protein EscR, affects the attachment of EDL933, suggesting a better basis for future studies of T3SS.

Prokaryotic expression, purification, identification, and polyclonal antibody prepa-ration of enterohemorrhagic Escherichia coli effector NleB1 / 军事医学

Objective To construct a prokaryotic plasmid expressing the recombinant protein of enterohemorrhagic Escherichia coli(EHEC) effector NleB1 and to prepare the polyclonal antibody of mouse anti-NleB1.Methods The nleB1 (990 bp) gene was amplified from the genome EHEC O157∶H7 and cloned into the expression plasmid pET24a to construct the recombinant plasmid pET24a-nleB1 that was transformed into E.coli BL21(DE3).After induction with isopropylthio-gelactoside( IPTG) , the His-tag fusion proteins were purified by Ni+affinity chromatography and gel slices.The polyclonal antibody was prepared by immunizing BALB/c mice with purified recombinant proteins and analyzed by Western blotting and ELISA.Results The pET24a-nleB1 recombinant plasmid was successfully constructed, the fusion protein was ex-pressed and purified,and the polyclonal antibody was obtained by immunizing mice with purified fusion protein.Western blotting and ELISA staining demonstrated that the polyclonal antibody was successfully obtained.Conclusion The prepara-tion of the polyclonal antibody against EHEC O157∶H7 NleB1 will be of help for further studies on the function of NleB1 protein.

Direct activation of expression vector withβ-casein gene promoter using engineered TALE transcription factors in mouse fibroblasts / 中国实验动物学报

Objective TALE-TFs were adopted to provide a new way in detection of the expression result ofβ-ca-sein gene promoter-interesting gene expression cassettes in mouse fibroblasts.Methods TALE-TFs of eukaryotic expres-sion plasmid and expression cassette withβ-casein gene promoter and red fluorescent protein reporter gene were co-nucleo-fected into mouse fibroblasts by Amaxa nucleofector.Results and Conclusion β-casein gene promoter was activated by artificial TALE-TFs in the mouse fibroblasts.The way is a new expression verification system instead of mammary epithelial cells with fibroblasts.

Self-made ultrasound/fluorescent bi-functional contrast agent for rabbit's normal lymph node imaging / 中国医学科学院学报

<p><b>OBJECTIVE</b>To prepare a lymph node-targeted ultrasound/fluorescence bi-functional imaging contrast agents, and observe its effectiveness both on contrast-enhanced ultrasound (CEUS) and vivo near infrared fluorescence (NIR) imaging through animal experiments.</p><p><b>METHODS</b>The chimeric lymph node-targeted ligand (phosphatidylserine) and near-infrared fluorescent substance were assembled to form bi-functional contrast microbubbles. The morphology and size distribution were detected by optical microscope and Malvern potential tests. Five normal New Zealand white rabbits were subcutaneously injected with the prepared contrast agent in bilateral footpads, and the imaging effectiveness of lymph nodes and lymphatic vessel were observed by CEUS and NIR technique. Then blue dye was subcutaneously injected at the same site, and the rabbits were sacrificed for lymph nodes pathological examination.</p><p><b>RESULTS</b>Lipid ultrasound microbubbles,with a mean size of 3-5 Μm in diameter, appeared to be uniform in distribution and regular in configuration. The images of inflow lymphatic vessel and relevant lymph node were quickly showed up after the subcutaneous injection by CEUS, which was identical to the result detected by NIR. Biopsy confirmed that all the blue-stained lymph nodes could be displayed by NIR.</p><p><b>CONCLUSIONS</b>The self-made bi-functional contrast agent has a good imaging ability in CEUS and NIR imaging. It may be a better agent as lymph node tracer.</p>

Repair of lower extremity traumatic soft tissue defect with ALT polyfoliate perforator flap / 中华整形外科杂志

<p><b>OBJECTIVE</b>Application of anterior lateral thigh flap (ALT) polyfoliate perforator flap for lower extremity soft tissue defect.</p><p><b>METHODS</b>From 2009 to 2011, anteral thigh perforator flap free transplantation were applied for traumatic soft tissue deformity of body surface.</p><p><b>RESULTS</b>There were 21 cases in this clinical application and the necrosis in distal part of two flaps were treated by free grafting. The other 19 flaps survived completely and the contour and function were satisfied.</p><p><b>CONCLUSION</b>ALT polyfoliate perforator flap could repair the complicated body surface soft tissue defect economically.</p>