Bacterial therapy has attracted increasing attention due to its special mechanism and abundant applications. With the flourishing development of synthetic biology, therapeutic genes have been introduced into engineering bacteria to improve their antitumor efficacy. However, it is difficult to spatiotemporally control the expression of these therapeutic genes at the tumor site in vivo, thereby considerably limiting the application of engineered bacteria in tumor treatment. To resolve this problem, we constructed a temperature-responsive bacterial strain capable of triggering the expression of exogenous genes in a laser-controllable way. Noxa, a pro-apoptotic protein, is chosen to test the expression of exogenous protein and its anti-tumor effect in engineered bacteria upon laser irradiation. Firstly, Noxa was fused to the C-terminus of the bacterial outer membrane protein cytolysin A (ClyA), and then the recombinant gene fragment ClyA-Noxa was inserted into the temperature-sensitive plasmid pBV220 and the recombinant plasmid was transformed into non-pathogenic Escherichia coli MG1655. Thus, we constructed the engineering strain (TRB@Noxa) that could express Noxa on the bacterial surface. TRB@Noxa could target and colonize the tumor tissue without causing notable host toxicity. The bacterial infection triggered thrombosis in the tumor tissue, resulting in the darkness of tumor sites. In a xenograft mouse tumor model, our strategy demonstrated precise tumor targeting and strong tumor inhibition. In conclusion, we successfully constructed a new engineering bacterial strain TRB@Noxa. TRB@Noxa combined with photothermal therapy could arrest tumor growth in the absence of photosensitizers, which represents an appealing method for antitumor therapy in the future.
Escherichia coli/radiation effects* ; Animals ; Humans ; Lasers ; Mice ; Proto-Oncogene Proteins c-bcl-2/biosynthesis* ; Neoplasms/therapy* ; Genetic Engineering ; Cell Line, Tumor ; Escherichia coli Proteins/genetics*

Objective:To systematically review the real experience of type 1 diabetes in adolescents.Methods:The qualitative research on the experience of type 1 diabetes in adolescents was electronically searched in PubMed, Web of Science, Embase, EBSCO, Cochrane Library, China Biology Medicine disc, WanFang Database, China National Knowledge Infrastructure, and VIP. The search period was from database establishment to July 2023. Two evidence-based trained researchers independently conducted literature screening and data extraction, and the results were integrated using the Meta-synthesis.Results:A total of 13 articles were included, 42 results were extracted and classified into 11 categories, which were integrated into four results, namely disease perception, complex psychological experiences, disease management, and post-traumatic growth.Conclusions:The awareness and psychological experience of type 1 diabetes in adolescents should be paid attention to. Medical institutions, families and schools should provide all-round support for adolescents with type 1 diabetes to help them enhance their disease self-management, improve their health outcomes and promote their post-traumatic growth.

Objective:To evaluate psychometric properties of infertility related stress assessment instruments and the methodological quality of research.Methods:Research on the psychometric property evaluation of infertility related stress assessment instruments was electronically searched on China National Knowledge Infrastructure, China Biology medicine disc, WanFang Data, VIP, PubMed, Web of Science, Embase, Scopus, and CINHAL. The search period was from database establishment to November 1, 2023. Two researchers independently screened literature and extracted data, evaluated the included assessment tools using the Consensus-based Standards for the Selection of Health Measurement Instruments (COSMIN) guideline, and formed recommendations.Results:A total of 20 studies were included, including 8 assessment instruments of the Fertility Problem Stress Inventory, Infertility Distress Scale, Fertility Problem Inventory, Fertility Problem Inventory-Short Form, Copenhagen Multi-Centre Psychosocial Infertility-Fertility Problem Stress Scales, Copenhagen Multi-Centre Psychosocial Infertility-Fertility Problem Stress Scales-Short Form, Infertility-Related Stress Scale, and the Symptom and Stress Inventory for Infertility. The content validity of assessment instrument was not reported or uncertain, and the quality of evidence for the assessment tools was moderate, all of which were B-level recommendations.Conclusions:The Fertility Problem Inventory, Copenhagen Multi-Centre Psychosocial Infertility-Fertility Problem Stress Scales, Copenhagen Multi-Centre Psychosocial Infertility-Fertility Problem Stress Scales-Short Form and the Infertility-Related Stress Scale can be temporarily recommended for use, but the psychometric property of each tool still needs to be fully verified. Clinical medical and nursing staff need to choose appropriate assessment instruments based on actual situation.

AIM:To investigate the effect of ankyrin repeat domain 49(ANKRD49)on epithelial-mesenchy-mal transition(EMT)in NCI-H1299 cells,and to explore its mechanism.METHODS:The ANKRD49 was over-ex-pressed in NCI-H1299 cells.The morphological changes of ANKRD49-overpressing NCI-H1299 cells were observed under microscope.The mRNA and protein expression levels of EMT-related markers[E-cadherin,transforming growth factor-β1(TGF-β1),vimentin and α-smooth muscle actin(α-SMA)]and EMT-related transcription factors(Snail1,Slug,Twist and ZEB1)were detected by RT-qPCR Western blot.Immunofluorescence staining was performed to observe the localiza-tion and expression of E-cadherin and vimentin in ANKRD49-overexpressing cells or control cells.Immunohistochemical method was performed to examine the levels of E-cadherin,α-SMA,Snail,Slug and ZEB1 in lung tissues of nude mice in-oculated with ANKRD49-overexpressing H1299 cells or control cells.RESULTS:Compared with the control group,the ANKRD49 overexpressing cells showed mesenchymal cell morphology(fusiform and less tight connections).RT-qPCR and Western blot results showed that the mRNA and protein levels of mesenchymal markers vimentin and α-SMA in ANKRD49 overexpressing cells were significantly higher than those in cells of control group,while the mRNA and protein levels of epithelial marker E-cadherin were lower than those in cells of control group.Compared with control group,the im-munofluorescence intensity of E-cadherin of H1299 cells decreased in after ANKRD49 overexpression,while that of vimen-tin increased significantly.Snail,Slug and ZEB1 expression were significantly elevated in ANKRD49 overexpressing cells compared with control group.The levels of E-cadherin in lung tissues of nude mice inoculated with ANKRD49-overexpressing H1299 cells declined,while the levels of α-SMA,Snail,Slug and ZEB1 increased compared with those in control mice.CONCLUSION:ANKRD49 promoted EMT of NCI-H1299 cells by increasing the expression of Snail1,Slug and ZEB1 and consequent downreguation of E-cadherin and upregulation of vimentin and α-SMA.

Objective To study the effect of ankyrin repeat domain 49(ANKRD49)on the migration of human lung adenocarcinoma cell line NCI-H1299 and its mechanism.Methods NCI-H1299 cells were infected with lentivirus vector carrying ANKRD49 gene and shRNA targeting ANKRD49 to construct the cell models stably overexpressing and knocking down ANKRD49. Meanwhile,the control cell models infected with empty lentivirus vector and lentivirus vector with scramble sequences were constructed respectively. The expression levels of ANKRD49 mRNA and protein were detected by real-time fluorescence quantitative PCR and Western blot. The effect of ANKRD49 on cell migration was measured by scratch test. The mRNA and protein levels of matrix metalloproteinase(MMP)-2/9 and tissue inhibitor of metalloproteinase(TIMP)-1/2 were detected by real-time fluorescence quantitative PCR and Western blot. The protein expression levels of p65,p-p65,IκBα and p-IκBα were detected by Western blot.Results The levels of ANKRD49 mRNA and protein in the ANKRD49 overexpression group were significantly higher than those in the control group(t = 70. 02 and 45. 68,respectively,each P < 0. 001). Compared with the control group,the migration ability of cells in the ANKRD49 overexpression group significantly increased at 24 h and 48 h(t = 5. 343 and 3. 282,P = 0. 005 9 and 0. 030 4,respectively);The mRNA transcription levels and protein expression levels of MMP-2 and MMP-9 significantly increased(t = 9. 304 and 6. 193,P =0. 000 7 and 0. 003 5,respectively),while the mRNA and protein expression of TIMP-1 and TIMP-2 decreased significantly(t = 3. 858 and 3. 517,P = 0. 018 2 and 0. 024 5,respectively),and the values of MMP-2/TIMP-1 and MMP-9/TIMP-2 significantly increased(t = 17. 7 and 9. 682,P < 0. 001 and < 0. 01,respectively);The expression of p-p65 and pIκBα significantly increased,the total protein levels of p65 and IκBα showed no obvious change,and the values of p-p65/p65 and p-IκBα/IκBα significantly increased(t = 3. 962 and 5. 370,P = 0. 016 7 and 0. 005 8,respectively). However,knocking down of ANKRD49 presented the opposite results.Conclusion ANKRD49 promotes the migration of NCI-H1299cells by enhan-cing the expression of MMP-2/9,the values of MMP-9/TIMP-1 and MMP-2/TIMP-2 via activating NF-κB/p65 signa-ling pathway.

BACKGROUND:As a dominant breed pig in southwest China,the southern Yunnan small-ear pig has been widely used as an experimental animal in the basic research of other disciplines,but there are still no reports on its application in anterior cruciate ligament reconstruction. OBJECTIVE:To establish a large animal model of the southern Yunnan small-ear pig with anterior cruciate ligament with autologous Achilles tendon was established. METHODS:Twenty adult female Yunnan small-ear pigs were equally randomized into two groups.In the autologous Achilles tendon group,the right knee anterior cruciate ligament was reconstructed with autologous Achilles tendon as a graft,while in the sham-operated group,a similar operation was performed on the right knee without any treatment of the anterior cruciate ligament.General conditions of each pig were observed and recorded before and 12 months after surgery.Ligaments and grafts were taken for gross observation and MAS scoring.Hematoxylin-eosin staining was performed to observe morphological characteristics of ligaments.The staining and arrangement of type I and type Ⅲ collagen were evaluated by immunohistochemistry.Transmission electron microscopy was used to observe the type,size,diameter,ratio,and distribution of collagen fibers in ligaments. RESULTS AND CONCLUSION:All animals had normal diet and activity,good wound healing,no obvious inflammatory reaction,no local purulent infection,and no significant changes in mental and urinary conditions compared with those before surgery.The reconstructed cruciate ligament of the knee was intact,with no stiffness and normal range of motion.Both the anterior drawer and Lachman tests were negative.Gross observation of the graft:12 months after surgery,the grafts was in good position,with good integrity,obvious tension,ligament color close to the original anterior cruciate ligament,and complete surface synovial coverage.Most of the intraarticular ligaments in the autologous Achilles tendon group were defined as MAS I type and a few were defined as MAS Ⅱ type.In the sham-operated group,the intraarticular ligament was defined as MAS I type.Hematoxylin-eosin staining indicated that,12 months after surgery,collagen fibers in the autologous Achilles tendon group began to appear bundled,isotropic,and uniformly arranged,with more obvious isotropic corrugations,and the nuclei were mainly linear or spindle-shaped,which were similar to those in normal anterior cruciate ligament tissue of the sham-operated group.Immunohistochemistry results indicated that,12 months after surgery,there was a higher expression of type I collagen and significantly less expression of type Ⅲ collagen in the reconstructed anterior cruciate ligament in the autologous Achilles tendon group.The degree of type I and type Ⅲ staining was similar in the two groups.Under the transmission electron microscope,the diameter,arrangement and density of collagen fibers in the reconstructed anterior cruciate ligament of the autologous Achilles tendon group were similar to those of the original anterior cruciate ligament at 12 months after surgery,indicating that the ligament remodeling process had been basically completed in the autologous Achilles tendon group at 12 months after surgery.Through a comprehensive evaluation of animal general conditions,ligament general view,MAS score,hematoxylin-eosin staining,immunohistochemistry,and transmission electron microscopy observation,we successfully established a large animal model of anterior cruciate ligament reconstruction using autogenous Achilles tendon in southern Yunnan small-ear pigs,with good morphological,histological and ultrastructural results.

Objective:To analyze the genomic epidemiological subtyping of carbapenem resistant Klebsiella pneumoniae (CRKP) isolated from a Third-class A hospital in Zhengzhou. Methods:From December 4, 2019 to January 10, 2020, 67 strains of CRKP were isolated from the samples submitted by the clinical departments of a Third-class A teaching hospital in Zhengzhou for microbiological testing. Multi-locus sequence typing (MLST) and carbapenem resistance genes were identified by whole genome sequencing and sequence analysis. Based on the whole genome SNP, the phylogenetic tree was constructed, and 67 CRKP strains were divided into clonal groups. The isolation ward and date of each clone group were analyzed.Results:Sixty-seven CRKP strains were classified into four MLST types (STs), of which 64 were ST11. There were 62 ST11 strains carrying blaKPC-2 gene. Based on genome-wide SNP phylogenetic tree, 64 ST11 strains were divided into four clone groups, two of which were dominant clone groups, including 33 and 27 strains respectively; the other two clone groups only contained 2 strains respectively. There was no aggregation of the dominant clones in the isolation department and date. Conclusion:Multiple clonal groups of ST11 strain carrying blaKPC-2 gene are differentiated during spreading, and they can spread in parallel and independently in the same hospital.

Objective:To analyze the genomic epidemiological subtyping of carbapenem resistant Klebsiella pneumoniae (CRKP) isolated from a Third-class A hospital in Zhengzhou. Methods:From December 4, 2019 to January 10, 2020, 67 strains of CRKP were isolated from the samples submitted by the clinical departments of a Third-class A teaching hospital in Zhengzhou for microbiological testing. Multi-locus sequence typing (MLST) and carbapenem resistance genes were identified by whole genome sequencing and sequence analysis. Based on the whole genome SNP, the phylogenetic tree was constructed, and 67 CRKP strains were divided into clonal groups. The isolation ward and date of each clone group were analyzed.Results:Sixty-seven CRKP strains were classified into four MLST types (STs), of which 64 were ST11. There were 62 ST11 strains carrying blaKPC-2 gene. Based on genome-wide SNP phylogenetic tree, 64 ST11 strains were divided into four clone groups, two of which were dominant clone groups, including 33 and 27 strains respectively; the other two clone groups only contained 2 strains respectively. There was no aggregation of the dominant clones in the isolation department and date. Conclusion:Multiple clonal groups of ST11 strain carrying blaKPC-2 gene are differentiated during spreading, and they can spread in parallel and independently in the same hospital.

Objective To evaluate hepatic perfusion assessed by contrast-enhanced ultrasonography (CEUS) for predicting cirrhosis accurately and non-invasively.Methods Forty patients with cirrhosis and twenty-five healthy controls were given CEUS examination,and time-intensity curves were drawn as the regions of interest located in liver parenchyma by using QLAB analyzing soft.The parameters of the two groups as follows:intensity of arterial perfusion (Iap),intensity of total perfusion of liver parenchyma (Ipeak),intensity of portal venous perfusion (Ipp),the ratio of portal venous perfusion and total perfusion (Ipp/Ipeak) were compared by independent-samples t test,and the diagnostic value of parameters were analyzed by receiver operating characteristic (ROC) curve.Resnlts Iap was bigger,while Ipp,Ipp/Ipeak were smaller in patients than that in controls( P ＜0.001 ).But there was no significant difference on Ipeak between the two groups.When Iap,Ipp,Ipp/Ipeak were used for the diagnosis of cirrhosis,the sensitivity were 67.3 ％,92.7％,96.4％ and the specificity were 80.0％,96.0％,92.3％,respectively.Conclusions CEUS can reflect the changes of the blood perfusion of cirrhotic liver.CEUS parameters Iap,Ipp,Ipp/Ipeak are significant different between the two groups and can be the non-invasive diagosis parameters of cirrhosis.

Food Testing was the experimental and applied basic courses for Prevent Medicine.According as the problems in teaching,some advice on teaching content,teaching method,and laboratory construction and management was discussed in order to cultivate the innovative specialties and develop the students' experimental ability.