To construct a recombinant attenuated Salmonella strain carrying the p57 and BLID genes and to evaluate its inhibitory effect on breast cancer cells and apoptosis induction in vitro,the recombinant attenuated Salmonella LH430/pEGFP-p57-BLID was constructed.RT-PCR and Western blot were used to detect the transcription and expression of the recombinant attenuated Salmonella in cancer cells.The CCK-8 method was used to assess the safety of the recombinant at-tenuated Salmonella and its impact on the activity of breast cancer cells.The cell scratch assay was used to analyze the influence of the recombinant attenuated Salmonella on the migratory ability of cancer cells.Flow cytometry and Western blot were used to analyze the effect of the recombinant attenuated Salmonella on the apoptosis of MDA-MB-231 cells.The bands of target genes carried by the recombinant attenuated Salmonella could be amplified by PCR,and these target genes could be transcribed and expressed in MDA-MB-231 cells.The recombinant attenuated Salmonella showed good growth characteristics and genetic stability.Compared with the PBS group,recombi-nant attenuated Salmonella LH430/pEGFP-p57-BLID significantly reduced(P＜0.01)the activity of MDA-MB-231 cells and significantly inhibited(P＜0.01)the migration of MDA-MB-231 cells.Recombinant attenuated Salmonella LH430/pEGFP-p57-BLID can obviously increase the number of apoptosis in MDA-MB-231 cells,significantly upregulate(P＜0.01)the expression of apoptotic protein Bax,and significantly downregulate(P＜0.01)the expression of apoptotic protein Bcl-2.The results showed that the recombinant attenuated Salmonella had an inhibitory effect on breast cancer MDA-MB-231 cells and could induce apoptosis,which laid a foundation for the subsequent study of anti-tumor in vivo.

To construct a recombinant attenuated Salmonella strain carrying the p57 and BLID genes and to evaluate its inhibitory effect on breast cancer cells and apoptosis induction in vitro,the recombinant attenuated Salmonella LH430/pEGFP-p57-BLID was constructed.RT-PCR and Western blot were used to detect the transcription and expression of the recombinant attenuated Salmonella in cancer cells.The CCK-8 method was used to assess the safety of the recombinant at-tenuated Salmonella and its impact on the activity of breast cancer cells.The cell scratch assay was used to analyze the influence of the recombinant attenuated Salmonella on the migratory ability of cancer cells.Flow cytometry and Western blot were used to analyze the effect of the recombinant attenuated Salmonella on the apoptosis of MDA-MB-231 cells.The bands of target genes carried by the recombinant attenuated Salmonella could be amplified by PCR,and these target genes could be transcribed and expressed in MDA-MB-231 cells.The recombinant attenuated Salmonella showed good growth characteristics and genetic stability.Compared with the PBS group,recombi-nant attenuated Salmonella LH430/pEGFP-p57-BLID significantly reduced(P＜0.01)the activity of MDA-MB-231 cells and significantly inhibited(P＜0.01)the migration of MDA-MB-231 cells.Recombinant attenuated Salmonella LH430/pEGFP-p57-BLID can obviously increase the number of apoptosis in MDA-MB-231 cells,significantly upregulate(P＜0.01)the expression of apoptotic protein Bax,and significantly downregulate(P＜0.01)the expression of apoptotic protein Bcl-2.The results showed that the recombinant attenuated Salmonella had an inhibitory effect on breast cancer MDA-MB-231 cells and could induce apoptosis,which laid a foundation for the subsequent study of anti-tumor in vivo.

Objective:To retrospectively analyze the independent risk factors of complicated appendicitis(CA)in children under five years old and establish a clinical prediction model, and to evaluate the clinical application of this model.Methods:A retrospective analysis was performed on children under five years old who underwent appendectomy at Children′s Hospital of Shanghai Jiao Tong University School of Medicine from January 2018 to December 2021.The children were divided into CA group and uncomplicated appendicitis group according to whether there was sign of perforation or gangrene in appendiceal tissue after operation.The differences in clinical features and preoperative laboratory test results between two groups were compared.The independent risk factors of CA were identified and a clinical prediction model was established.The clinical prediction model was verified by receiver operating characteristic curve.Results:A total of 140 children were enrolled in this study, including 84 cases in the CA group and 56 cases in uncomplicated appendicitis group.Univariate and binary Logistic regression analysis showed that the duration of symptoms>23.5 h( OR=6.650, 95% CI 2.469-17.912, P<0.05), abdominal muscle tension( OR=3.082, 95% CI 1.190-7.979, P<0.05) and C-reactive protein>41 mg/L ( OR=3.287, 95% CI 1.274-8.480, P<0.05) were independent risk factors for CA( P<0.05). The clinical prediction model of CA was constructed by the above mentioned three independent risk factors.The area under the receiver operating characteristic curve of the clinical prediction model was 0.881(95% CI 0.825-0.936), the sensitivity was 77.4%, the specificity was 87.5%, the positive predictive value was 91.3% and the negative predictive value was 70.0%. Conclusion:Acute appendicitis in children under five years old is more likely to progress to CA if the duration of symptoms>23.5 h, the level of C-reactive protein is increased, and the abdominal muscle tension is accompanied.The clinical prediction model of CA constructed by common clinical information in pediatric clinics has good prediction efficiency, which provides a simple and feasible reference method for clinicians to distinguish CA from uncomplicated appendicitis.

ObjectiveTo investigate the pharmacodynamic characteristics and explore the molecular mechanism of Honghua oral liquid （HOL） in relieving neuropathic pain （NP）. MethodHealthy male SD rats were randomly assigned into sham group， model group， low-， medium-， high-dose （0.5， 1.0， 2.0 mL·kg^-1·d^-1， respectively） HOL groups， and a positive drug （pregabalin， 25 mg·kg^-1·d^-1） group， with 6 rats in each group. Spinal nerve ligation （SNL） of L5 was conducted in other groups except the sham group. Drug administration was performed 3 days after the SNL surgery for 2 consecutive weeks， and samples were collected after the end of the administration. During the treatment period， the mechanical pain threshold and cold pain threshold were determined to measure the pain-relieving effect of HOL. Transcriptome sequencing was performed on hippocampal tissue samples from the sham， model， and high-dose HOL groups， and differentially expressed genes between the sham group and the model group as well as the model group and HOL high-dose group were obtained. After pathway enrichment analysis， we selected the targets which were closely related to neuroinflammation for validation， and predicted the specific binding sites of the major active components in HOL with the targets through molecular docking. In addition， the serum levels of tumor necrosis factor-α （TNF-α） and interleukin-10 （IL-10） were determined by enzyme-linked immunosorbent assay （ELISA） to evaluate the effect of HOL on neuroinflammation in NP rats. ResultCompared with the sham group， SNL decreased the mechanical pain threshold and cold pain threshold （P<0.05）. Compared with the model group， HOL recovered the mechanical pain threshold and cold pain threshold （P<0.05）. The transcriptome data showed that 376 differentially expressed genes （DEGs） were identified between the model group and the sham group， including 124 upregulated genes and 252 downregulated genes， and 194 DEGs between the model group and the high-dose HOL group， including 33 upregulated genes and 161 downregulated genes. Among them， insulin-like growth factor 1（IGF1）， matrix metallopeptidase-2 （MMP-2）， matrix metallopeptidase-14 （MMP-14）， erb-B2 receptor tyrosine kinase 2 （ERBB2）， and integrin subunit alpha 5 （ITGA5） associated with NP were selected for further validation. The Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） results showed that compared with the sham group， the modeling up-gurelated the mRNA levels of the above five molecules in the hippocampus （P<0.01）. Compared with model group， HOL down-regulated the mRNA levels of these molecules （P<0.01）. The molecular docking results showed that the main active components of safflower， hydroxysafflor yellow A， kaempferol， and quercetin， formed stable hydrogen bonds with the amino acid residues of IGF1， MMP-2， MMP-14， ERBB2， and ITGA5. The enzyme-linked immunosorbent assay（ELISA） results showed that compared with those in the sham group， the serum levels of TNF-α and IL-10 were out of balance in the model rats （P<0.01）. Compared with the model group， HOL lowered the level of the pro-inflammatory cytokine TNF-α （P<0.01） and elevated that of the anti-inflammatory cytokine IL-10 （P<0.05）. ConclusionHOL exerts analgesic effect on SNL rats by inhibiting neuroinflammation.

Objective:To identify core contents of health education for patients with pemphigus by using the Delphi method.Methods:On the basis of literature review and group discussions, two rounds of questionnaire survey were performed in 17 experts to evaluate the importance of health education items for patients with pemphigus by using the Delphi method. Indices including importance score, coefficient of variation, authority coefficient and Kendall coefficient of concordance were calculated to identify core contents of health education for patients with pemphigus.Results:In the two rounds of questionnaire survey, all experts complete the questionnaires, the coefficients of variation were all less than 0.35, the authority coefficient was 0.893, and the Kendall coefficients of concordance were 0.283 and 0.288 respectively ( χ2= 168.17, 181.25, respectively, both P < 0.001) . After the two rounds of expert consultation, the core contents of health education for patients with pemphigus were finally confirmed, which consisted of 6 primary items and 32 secondary items, and the primary items included "disease knowledge", "medication guidance", " dietary guidance", "skin care", "rest, activities and psychological guidance" and " follow-up and medical guidance". Conclusion:By using the Delphi method, the core contents of health education have been successfully identified for patients with pemphigus, which provide a basis for systematization and scientization of health education contents for patients with pemphigus.

d-α-Tocopheryl polyethylene glycol 1000 succinate (TPGS, also known as vitamin E-TPGS) is a biodegradable amphiphilic polymer prepared by esterification of vitamin E with polyethylene glycol (PEG) 1000. It is approved by the US Food and Drug Administration (FDA) and has found wide application in nanocarrier drug delivery systems (NDDS). Fully characterizing the in vivo fate and pharmacokinetic behavior of TPGS is important to promote the further development of TPGS-based NDDS. However, to date, a bioassay for the simultaneous quantitation of TPGS and its metabolite, PEG1000, has not been reported. In the present study, we developed such an innovative bioassay and used it to investigate the pharmacokinetics, tissue distribution and excretion of TPGS and PEG1000 in rat after oral and intravenous dosing. In addition, we evaluated the interaction of TPGS with cytochromes P450 (CYP450s) in human liver microsomes. The results show that TPGS is poorly absorbed after oral administration with very low bioavailability and that, after intravenous administration, TPGS and PEG1000 are mainly distributed to the spleen, liver, lung and kidney before both being slowly eliminated in urine and feces as PEG1000. In vitro studies show the inhibition of human CYP450 enzymes by TPGS is limited to a weak inhibition of CYP3A4. Overall, our results provide a clear picture of the in vivo fate of TPGS which will be useful in evaluating the safety of TPGS-based NDDS in clinical use and in promoting their further development.

To explore the effects of intrauterine infection on early growth and neurobehavioral development in neonatal rats. (E. coli) was inoculated into uterine cervix of pregnant rats with gestation of 15 d to establish the intrauterine infection model, and the effect on the delivery of pregnant rats was observed. The neonatal rat brain tissue was stained with Hematoxylin-Eosin and the cerebral white matter damage was assessed. Immunohistochemical staining and Western blot analysis were performed to evaluate the expression of glial fibrillary acidic protein (GFAP), 2', 3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) and neurofilament (NF) in pup brains. Birth weight and early growth development indices were monitored,and neurobehavioral tests were performed to access the change of neurobehavioral development in neonatal rats. The white blood cell count increased significantly in the uterus and placenta of the pregnant rats after intrauterine E. coli infection and no significant impact was observed on the delivery of pregnant rats. Weak staining and focal rarefaction of cerebral white matter from rats at P7 in intrauterine infection group were observed. The expression of GFAP markedly increased (<0.05) in infection group, while the level of CNPase and NF in pup brains at P7 significantly decreased (<0.05 or <0.01). Compared with control group, the neonatal rats in infection group had lower birth weight and slower weight gain during the suckling period (<0.05 or <0.01), and the completion times of ear opening, eye opening, surface righting, negative geotaxis, acoustic startle and swimming test in infection group were significantly delayed (<0.05 or <0.01). Intrauterine infection in pregnant rats can induce cerebral white matter damage and retardation of early growth and neurobehavioral development in neonatal rats.
Animals ; Animals, Newborn ; Behavior, Animal ; Body Weight ; Disease Models, Animal ; Escherichia coli ; Escherichia coli Infections ; complications ; physiopathology ; Female ; Glial Fibrillary Acidic Protein ; genetics ; Growth Disorders ; etiology ; Leukoencephalopathies ; etiology ; Pregnancy ; Pregnancy Complications, Infectious ; physiopathology ; Rats ; Rats, Sprague-Dawley

Intravascular ultrasound (IVUS) is widely used in coronary artery examination. Ultrasonic elastography combined with IVUS is very conspicuous in identifying plaque component and in detecting plaque vulnerability degree. In this study, a simulation model of the blood vessel based on finite element analysis (FEA) was established. The vessel walls generally have radial changes caused by different intravascular pressure. The signals at lower pressures were used as the pre-deformation data and the signals at higher pressure were used as the post-deformation data. Displacement distribution was constructed using the time-domain cross-correlation method, and then strain images. By comparison of elastograms under different pressures, we obtained the optimal pressure step. Furthermore, on the basis of the obtained optimize pressure step, the simulation results showed that this method could effectively distinguish characteristics between different component plaques, and could guide the later experiments and clinical applications.
Angiography ; Arteries ; diagnostic imaging ; pathology ; Elasticity Imaging Techniques ; Finite Element Analysis ; Humans ; Plaque, Atherosclerotic ; diagnostic imaging ; Pressure

Intraoperative analysis of the position of microelectrode in the globus pallidus only depends on experiences and subjective interpretation of microeletrode recording discharge of neurons during pallidotomy for Parkinson's disease. In this paper is reported a method for objective localization of microeletrode during neurosurgery. This method uses the factor of interspike interval based on microelectrode recordings. It is effective for identifying the change of the globus pallidal organism along the microelectrode pathway and the departure or deviation of the needle-pathway. The classification is consistent with the anatomic structures and the results of neurosurgery. This method can be used as a guide in pallidotomy. Globus pallidus.
Adult ; Aged ; Female ; Globus Pallidus ; surgery ; Humans ; Male ; Microelectrodes ; Middle Aged ; Pallidotomy ; methods ; Parkinson Disease ; surgery ; Stereotaxic Techniques ; Surgery, Computer-Assisted ; methods

0.05), but not in other brain regions. The number of GFAP-immunopositive cells of the E.coli-treated pups was markedly increased in periventricular white matter and hippocampus at P7 compared with the control group (P0.05). CONCLUSION: Intrauterine infection induces an increased expression of GFAP in the neonatal brain. [