[Clinical analysis of the correlation between gallbladder adenomyomatosis and occult pancreaticobiliary reflux].

Objective: To explore the association between gallbladder adenomyomatosis (GA) and occult pancreaticobiliary reflux (OPBR). Methods: A total of 81 patients with GA who underwent cholecystectomy in Shanghai East Hospital from December 2020 to January 2022 were enrolled, including 48 cases of fundal type, 28 cases of segmental type and 5 cases of diffuse type. Patient's intraoperative bile was coltected and tested for amylase. According to gallbladder bile amylase level, patients were divided into OPBR group (bile amylase>110 U/L) and the control group (bile amylase≤110 U/L). Results: Among 81 patients, 32 were male and 49 were female, and aged (49.1±13.2) years; there were 66 cases in control group, including 27 males and 39 females, and aged (50.0±12.9)years; there were 15 patients in the OPBR group, including 5 males and 10 females, and aged (45.1±14.2) years. In terms of the clinical features of the two groups, there was no significant difference (all P>0.05), except for a significant increase in biliary amylase in the OPBR group compared with the control group (P<0.001). However, the incidence of OPBR was significantly different in the three types of GA, with a lower incidence of OPBR in the fundal type (10.4%, 5/48) than in the segmental type (28.6%, 8/28) and diffuse type (2/5) (P=0.038). In addition, segmental GA was more likely to be combined with gallbladder stones (85.7%, 24/28) than fundal GA (58.3%, 28/48) and diffuse GA (3/5) (P=0.031). Univariate and multivariate logistic regression analyses showed OPBR [OR (95%CI)=3.410 (1.010 to 11.513), P=0.048] and combined gallbladder stones [OR (95%CI)=2.974 (1.011 to 8.745), P=0.048] indepenclently correlated with segmental and diffuse GA. Conclusions: The incidence of OPBR is higher in segmental and diffuse GA, and gallstones and OPBR are independently associated with the occurrence of segmental and diffuse GA. These results suggest that OPBR may be the initiating factor for the occurrence and carcinogenesis of segmental and diffuse GA.

CircABCB10 silencing inhibits the cell ferroptosis and apoptosis by regulating the miR-326/CCL5 axis in rectal cancer.

Circular ATP binding cassette subfamily B member 10 (circABCB10) has been identified to have oncological functions in several tumors. However, the roles of circABCB10 in rectal cancer remain unknown. The expression of circABCB10, microRNA (miR)-326 and C-C motif chemokine ligand 5 (CCL5), and apoptosis related-protein was detected using quantitative real-time polymerase chain reaction or western blot, respectively. Cell survival or apoptosis was measured using cell counting kit-8 assay or flow cytometry. The accumulations of intracellular lipid reactive oxygen species (ROS) and Fe2+ were analyzed using C11-BODIP dye or iron kit assay, respectively. In vivo experiments were conducted using the murine xenograft model. The interaction between miR-326 and circABCB10 or CCL5 was confirmed by dual-luciferase reporter assay and RNA immunoprecipitation assay. CircABCB10 and CCL5 were upregulated but miR-326 was downregulated in rectal cancer. The knockdown of circABCB10 promoted rectal cancer cell ferroptosis and apoptosis in vitro as well as inhibited tumor growth in vivo. miR-326 was a target of circABCB10, and the miR-326 inhibition could partially attenuate circABCB10 deletion-induced cell ferroptosis and apoptosis. miR-326 directly interacted with CCL5, and the miR-326 inhibition suppressed cell ferroptosis and apoptosis by targeting CCL5. Besides, we observed that miR-326 was negatively regulated by circABCB10, while CCL5 was positively regulated by it, and circABCB10 served as a sponge of miR-326 to regulate the CCL5 expression in rectal cancer cells. CircABCB10 silence promoted rectal cancer cell ferroptosis and apoptosis by regulating the miR-326/CCL5 axis, suggesting a potential therapeutic target for rectal cancer therapy.

Retraction: Olefin epoxidation with chiral salen Mn(iii) immobilized on ZnPS-PVPA upon alkyldiamine.

[This retracts the article DOI: 10.1039/C6RA00002A.].

Inactivation of Rab23 inhibits the invasion and motility of pancreatic duct adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) is charac-terized by a poor prognosis and high mortality rate. In this study, we investigated the expression of Rab23 in non-tumor pancreatic tissues and PDACs via immunohistochemistry. Rab23 was found in 39 of 58 (67.2%) and in 11 of 30 (36.7%) of the PDAC and non-tumor pan-creatic tissue samples (P = 0.0073), respectively. There were signifi-cant correlations between Rab23 expression and unfavorable variables, including cancer differentiation level (P = 0.0089), lymph nodal (P = 0.0099), and distant metastases (P = 0.0173). Inactivation with small interfering RNA against Rab23 in the human pancreatic cancer cell line Panc-1 inhibited the migration and invasive potential of the cells. Our data provide new insight into the essential role of Rab23 in PDAC inva-sion and metastasis and suggest that Rab23 expression is a useful indi-cator of metastatic potential; hence, it may be a new therapeutic target for this common malignancy.

Dynamic evolution and immunoreactivity of aptamers binding to polyclonal antibodies against MPT64 antigen of Mycobacterium tuberculosis.

Antibody responses can be useful markers of tuberculosis infection. However, the established immunoassay diagnostic method is limited by antigenic variability. Replacing the recombinant proteins with aptamers may overcome these antigenic challenges. In this study, we systematically monitored the selection process of aptamers against anti-MPT64 antibodies of Mycobacterium tuberculosis to obtain more aptamers for developing a multisite system to increase the sensitivity of TB serological diagnosis. Twelve high-affinity aptamers with distinctive secondary structures were obtained by analyzing the dynamic evolution of aptamers against anti-MPT64 antibodies in the process of system evolution of ligands by exponential enrichment (SELEX). Pocket and stem-loops were found to be the basis of these aptamers binding to antibodies. Point mutations of highly conserved nucleotides in the pocket and stem-loop structures resulted in decreased affinity of aptamers to targets. To test the potential of these aptamers for future use in a serological diagnostic tool, three high-affinity aptamers with different epitope specificities were applied as capture aptamer in an enzyme-linked immunosorbent assay (ELISA) with sera of TB patients. The results showed that three aptamers all effectively bound anti-MPT64 antibodies from TB patients and had high specificity and sensitivity. These aptamers with high immunoreactivity in human sera may represent an efficient and promising analogue of MPT64 and have potential to substitute MPT64 as a nucleic acid antigen in the serological diagnosis of TB. Moreover, these aptamers with different epitope specificities may facilitate the development of a sandwich assay platform or a multisite system to effectively capture more targets in sera.

A phase II study of sirolimus, tacrolimus and rabbit anti-thymocyte globulin as GVHD prophylaxis after unrelated-donor PBSC transplant.

We report on a prospective phase II trial of 32 patients who underwent unrelated-donor hematopoietic cell transplantation, with a tacrolimus, sirolimus and rabbit anti-thymoctye globulin GVHD prophylactic regimen. The primary study endpoint was incidence of grades II-IV acute (aGVHD), with 80% power to detect a 30% decrease compared with institutional historical controls. Median age at transplant was 60 (19-71). In total, 23 patients (72%) received reduced-intensity conditioning, whereas the remainder received full-intensity regimens. Median follow-up for surviving patients was 35 months (range: 21-49). The cumulative incidence of aGVHD was 37.3%, and the 2-year cumulative incidence of chronic GVHD was 63%. We observed thrombotic microangiopathy in seven patients (21.8%), one of whom also developed sinusoidal obstructive syndrome (SOS). Four of the 32 patients (12.5%) failed to engraft, and 3 of these 4 died. As a result, enrollment to this trial was closed before the targeted accrual of 60 patients. Two-year OS was 65.5% and EFS was 61.3%. Two-year cumulative incidence of relapse was 12.5% and non-relapse mortality (NRM) was 15.6%. NRM and aGVHD rates were lower than historical rates. However, the unexpectedly high incidence of graft failure requires caution in the design of future studies with this regimen.

Efficacy of recombinant adenovirus-mediated double suicide gene therapy in human keloid fibroblasts.

BACKGROUND: The recombinant adenovirus-mediated double suicide gene CDglyTK has been widely used for gene therapy of cancer. The biological behaviour of keloids is similar to that of cancer, in that they may extend beyond the site of injury, and do not subside. There are no effective strategies available for keloid therapy. Gene therapy is gaining greater importance in the field of plastic surgery, and the CDglyTK double suicide gene/prodrug system has been receiving greater attention. AIM: To show the lethal and bystander effects of a double suicide gene in keloid fibroblasts. Bcl-2 and BAX play an important role in the apoptosis induced by the double suicide gene. METHODS: Recombinant adenovirus expression CDglyTK suicide genes were constructed using the modified AdEasy system. The lethal and bystander effects were measured after 48 h using the MTT assay. The morphological changes in fibroblasts were detected by haematoxylin and eosin staining, and apoptosis was detected by the terminal dUTP nick-end labelling assay. Bcl-2 and BAX were detected by immunohistochemistry and quantitative real-time PCR. RESULTS: CDglyTK could be constructed easily and relatively quickly. The lethal and bystander effects of CDglyTK were marked in keloid fibroblasts. Apoptosis was one of the main processes leading to fibroblast death in keloids, and Bcl-2 and BAX played an important role in the process of apoptosis. CONCLUSION: Together, the results support the notion that recombinant adenovirus-mediated CDglyTK double suicide gene therapy is effective in destroying keloid fibroblasts and provide a sound scientific rationale for keloid trials in vivo.

[Cloning of a full length cDNA of human thrombopoietin receptor c-Mpl and construction of engineered cells that stably express c-mpl].

A full length cDNA fragment encoding for human thrombopoietin receptor c-Mpl has been amplified by RT-PCR from the total RNA of human HEL cells. The complete sequence of the cloned cDNA was determined and is identical to that previously reported. Then the fragment was subcloned into the mammalian expression vector pcDNA3 and the resulting plasmid is designated as pcMPL. K562 cells, which do not express c-mpl, were transfected with pcMPL and pcDNA3, respectively. The transformants were selected with G418 and then tested by Northern and Southern blotting. A group of engineered cell lines stably expressing c-mpl have been obtained, which will facilitate further research on the signaling mediated by c-Mpl.

[Effect of L1ECD on mouse primarily cultured neurons and construction of transgenic mice specifically expressing L1ECD in brain].

Neural cell adhesion molecule L1 is an important molecule mediating cell-cell interactions during the development of nervous system. L1 can promote axonal outgrowth and is related with nerve cell migration, and therefore L1 plays an important role both in the development and maintaince of the nervous system. In humans, mutations in the L1 gene can lead to mental retardation, spastic paraplegia, hydrocephalus, and other developmental abnormalities. The molecular mechanisms of mutations in L1 gene to induce inherited neurological diseases are not clear. In present investigation, a transgenic DNA of mouse L1 extracellular domain (L1ECD) was constructed by adding a stop codon to the end of L1ECD cDNA and then putting it under the control of CAMK II promoter, which is active specifically in the brain. To verify this construct, L1ECD cDNA was subcloned into an expression vector pCEP4 and then transfected the C6 cells. The expression of L1ECD cDNA in C6 cells was confirmed by Northern blotting and the effects of L1ECD on the growth rate and morphology of C6 cells in vitro as well as primarily cultured neurons were observed. The L1ECD constructs were microinjected into the fertilized zygotes of C57BL/6 mice. The transgenic mice thus produced were identified by Southern and Northern hybridization analysis. The results demonstrated that the L1ECD was integrated in the genome of transgenic mice and expressed specifically in the brain.