Retrospective assessment of short-term outcomes of robotic- versus laparoscopic-assisted duodenal diamond anastomosis in neonates.

OBJECTIVE: The purpose of this study was to retrospectively compare the short-term outcomes of robotic- (RAD) and laparoscopic-assisted duodenal diamond-shaped anastomosis (LAD) in neonates. METHODS: Neonates who underwent RAD (n = 30) or LAD (n = 38) between January 2019 and December 2022 were analyzed retrospectively. Major patient data were collected, including preoperative, intraoperative, and postoperative information. RESULTS: All patients were neonates below the age of 30 days weighing 4 kg. Thirty (44.1%) neonates underwent RAD and 38 neonates (55.9%) underwent LAD. Compared to the LAD group, the RAD group had a shorter intra-abdominal operation time (RAD, 60.0(50.0 ~ 70.0) min; LAD, 79.9(69.0 ~ 95.3) min; p < 0.001). There were no significant differences in immediate and 30-day complications between the two groups. CONCLUSIONS: RAD is safe and effective in neonates. Compared to traditional LAD, RAD showed comparable results.

Expression and possible role of Smad3 in postnecrotizing enterocolitis stricture.

Objective: To investigate the expression of Smad3 (mothers against decapentaplegic homolog 3) protein in postnecrotizing enterocolitis stricture and its possible mechanism of action. Methods: We used immunohistochemistry to detect the expression characteristics of Smad3 and nuclear factor kappa B (NF-κB) proteins in human postnecrotizing enterocolitis stricture. We cultured IEC-6 (crypt epithelial cells of rat small intestine) in vitro and inhibited the expression of Smad3 using siRNA technique. Quantitative PCR, western blotting, and ELISA were used to detect the changes in transforming growth factor-ß1 (TGF-ß1), NF-κB, tumor necrosis factor-α (TNF-α), vascular endothelial growth factor (VEGF), and zonula occludens-1 (ZO-1) messenger RNA (mRNA) and protein expressions in IEC-6 cells. CCK8 kit and Transwell cellular migration were used to detect cell proliferation and migration. Changes in epithelial-mesenchymal transition (EMT) markers (E-cadherin and vimentin) in IEC-6 cells were detected by immunofluorescence technique. Results: The results showed that Smad3 protein and NF-κB protein were overexpressed in narrow intestinal tissues and that Smad3 protein expression was positively correlated with NF-κB protein expression. After inhibiting the expression of Smad3 in IEC-6 cells, the mRNA expressions of NF-κB, TGF-ß1, ZO-1, and VEGF decreased, whereas the mRNA expression of TNF-α did not significantly change. TGF-ß1, NF-κB, and TNF-α protein expressions in IEC-6 cells decreased, whereas ZO-1 and intracellular VEGF protein expressions increased. IEC-6 cell proliferation and migration capacity decreased. There was no significant change in protein expression levels of EMT markers E-cadherin and vimentin and also extracellular VEGF protein expression. Conclusions: We suspect that the high expression of Smad3 protein in postnecrotizing enterocolitis stricture may promote the occurrence and development of secondary intestinal stenosis. The mechanism may be related to the regulation of TGF-ß1, NF-κB, TNF-α, ZO-1, and VEGF mRNA and protein expression. This may also be related to the ability of Smad3 to promote epithelial cell proliferation and migration.

Expression and Possible Role of Silent Mating Type Information Regulation 2 Homolog 1 in Post-necrotizing Enterocolitis Stricture in vivo and in vitro.

Purpose: To investigate the expression and possible role of Sirtuin1 or Silent mating-type information regulation 2 homolog-1 (SIRT1) in post-necrotizing enterocolitis stricture. Materials and Methods: The expression characteristics of SIRT1 and TGF-ß1 in post-necrotizing enterocolitis stricture were detected by immunohistochemistry. The siRNA-SIRT1 was used to inhibit the expression of SIRT1 in intestinal epithelial cells-6 (IEC-6), and qRT-PCR, WB, and ELISA were utilized to detect the changes of Transforming growth factor-ß1 (TGF-ß1), nuclear factor (NF)-κB, tumor necrosis factor-α (TNF-α), tight junction protein-1 (ZO-1), and vascular endothelial growth factor (VEGF) expressions. The IEC-6 cell proliferation and migration ability were tested via CCK8 kit and Transwell test. The expression of E-cadherin and Vimentin in cells was detected by immunofluorescence. Results: The CRP, IL-6, IL-10, and IFN-γ in the serum of Necrotizing enterocolitis (NEC) intestinal stenosis patients were significantly higher than the reference values. The SIRT1 protein was under-expressed and the TGF-ß1 protein was overexpressed in NEC intestinal stenosis tissue. And the expression of SIRT1 was negatively correlated with TGF-ß1. At the time of diagnosis of NEC, the expression of SIRT1 decreased in children with respiratory distress syndrome and CRP level increased. After inhibiting the expression of SIRT1 in IEC6 cells, the expression levels of TGF-ß1, Smad3, and NF-κB were decreased, and the expression of ZO-1 was also decreased. The proliferation and migration ability of IEC6 cells was decreased significantly, and the expression of E-cadherin and Vimentin proteins in IEC6 cells did not change significantly. Conclusion: Promotion of intestinal fibrosis by inflammation may be the mechanism of post-necrotizing enterocolitis stricture. SIRT1 may be a protective protein of NEC. The probable mechanism is that SIRT1 can regulate intestinal fibrosis and can protect the intestinal mucosal barrier function to participate in the process of post-necrotizing enterocolitis stricture.

A single-CRD C-type lectin from Haliotis discus hannai acts as pattern recognition receptor enhancing hemocytes opsonization.

C-type lectins (CTLs), as a member of the Ca2+-dependent carbohydrate recognition protein superfamily, play multiple roles in non-self recognition and the elimination of invading pathogens. In this study, a C-type lectin was identified and characterized from the Pacific abalone Haliotis discus hannai (designed as HdClec), and its open reading frame (ORF) encoded a polypeptide of 163 amino acids containing a typical signal peptide and only one carbohydrate-recognition domain (CRD). The deduced amino acid sequence of CRD in HdClec shared identities ranging from 22.4% to 39.8% with that of other identified CRDs of CTLs. A novel NPN motif was found in Ca2+-binding site 2 of HdClec. The mRNA transcripts of HdClec were detectable in all the examined tissues of non-stimulated abalones, with the highest expression in hepatopancreas (224.13-fold of that in gills). The expression of HdClec mRNA in hemocytes was significantly up-regulated after Vibrio harveyi challenge. Recombinant HdClec protein (rHdClec) could bind lipopolysaccharide (LPS) and peptidoglycan (PGN) in vitro in the presence of Ca2+. Coinciding with the PAMPs binding assay, rHdClec displayed broad agglutination activities towards Gram-negative bacteria V. splendidus, V. anguillarum, V. parahaemolyticus, V. harveyi, Escherichia coli, and Gram-positive bacteria Micrococcus luteus. Moreover, rHdClec could significantly elicit the chemotactic response of hemocytes in vitro. And the phagocytosis and encapsulation ability of hemocytes could be significantly enhanced by rHdClec. All these results showed that HdClec could function as pattern recognition receptors (PRRs) and further enhance the opsonization of hemocytes, which might play a crucial role in the innate immune responses of Pacific abalone.

Effects of the potential probiotic Bacillus subtilis D1-2 on growth, digestion, immunity and intestinal flora in juvenile sea cucumber, Apostichopus japonicus.

In the present study, a potential probiotic Bacillus subtilis D1-2 with antibacterial activity was isolated from the gut of Apostichopus japonicus. The purpose of this experiment was to assess the effect of B. subtilis D1-2 at different concentrations (C: 0 CFU/g, BL: 105 CFU/g, BM: 107 CFU/g and BH: 109 CFU/g) on the growth performance, digestive enzyme activity, immune ability and intestinal flora of A. japonicus. After the 56-day feeding trial, the final body weight and weight gain rate of juvenile sea cucumber A. japonicus fed B. subtilis D1-2 were significantly increased, especially in the BM group. Additionally, the lipase activity of the intestine was significantly increased in the BM and BH groups. Enhanced immunity was also found in sea cucumbers supplemented with B. subtilis D1-2. Alpha diversity indices showed that the B. subtilis D1-2-supplemented groups had higher intestinal microbial richness and diversity than the control group. The beta diversity analysis indicated that the bacterial communities in the B. subtilis D1-2-supplemented groups were quite similar but different from the bacterial communities in the control group. Dietary supplementation with B. subtilis D1-2 increased the relative abundance of some potential probiotic-related genera, including Lactobacillus, Clostridium, Lactococcus, Bifidobacterium and Streptococcus. In conclusion, dietary addition of B. subtilis D1-2 could effectively promote the growth of A. japonicus, improve its digestion and immunity capacity to a certain extent, and actively regulate the intestinal microflora of A. japonicus.

Antibacterial activities and mechanisms of action of a defensin from manila clam Ruditapes philippinarum.

Defensins represent an evolutionary ancient family of antimicrobial peptides, which played an undeniably important role in host defense. In the present study, a defensin isoform was identified and characterized from manila clam Ruditapes philippinarum (designed as Rpdef1α). Multiple alignments and phylogenetic analysis suggested that Rpdef1α belonged to the defensin family. Quantitative RT-PCR and immunohistochemical analysis revealed that Rpdef1α transcripts and the encoding peptide were dominantly expressed in the tissues of gills and mantle. After Vibrio anguillarum challenge, the Rpdef1α transcripts were significantly up-regulated in gills of clams. In addition, rRpdef1α not only showed broad-spectrum antimicrobial activities towards Vibrio species, but also inhibited the formation of bacterial biofilms. Knockdown of Rpdef1α transcripts caused significant increase in the cumulative mortality of manila clams post V. anguillarum challenge. Membrane integrity, scanning electron microscopy analysis and electrochemical assay indicated that rRpdef1α was capable of causing bacterial membrane permeabilization and then resulted in cell death. Moreover, phagocytosis and chemotactic ability of hemocytes could be significantly enhanced after incubation with rRpdef1α. Overall, these results suggested that Rpdef1α could act as both antibacterial agent and opsonin to defend against the invading microorganisms in manila clam R. philippinarum.

Maresin-1 Inhibits Oxidative Stress and Inflammation and Promotes Apoptosis in a Mouse Model of Caerulein-Induced Acute Pancreatitis.

BACKGROUND This study aimed to investigate the effects of maresin-1 (MaR1) in a mouse model of caerulein-induced acute pancreatitis (AP). MATERIAL AND METHODS Fifty C57BL/6 mice with caerulein-induced AP were divided into the untreated control group (N=10), the untreated AP model group (N=10), the MaR1-treated (low-dose, 0.1 µg) AP model group (N=10), the MaR1-treated (middle-dose, 0.5 µg) AP model group (N=10), and the MaR1-treated (high-dose, 1 µg) AP model group (N=10). Enzyme-linked immunoassay (ELISA) measured serum levels of amylase, lipase, tumor necrosis factor-alpha (TNF-alpha), interleukin-1ß (IL-1ß), and IL-6 and mRNA was measured by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Malondialdehyde (MDA), protein carbonyls, superoxide dismutase (SOD), and the ratio of reduced glutathione/oxidized glutathione (GSH/GSSG) were measured. Histology of the pancreas included measurement of acinar cell apoptosis using the terminal-deoxynucleotidyl transferase-mediated nick end-labeling (TUNEL) assay. Western blot measured Toll-like receptor 4 (TLR4), MyD88, and phospho-NF-kappaB p65, and apoptosis-associated proteins Bcl-2, Bax, cleaved caspase-3, and cleaved caspase-9. RESULTS Following treatment with MaR1, serum levels of amylase, lipase, TNF-alpha, IL-1ß, and IL-6 decreased, MDA and protein carbonyl levels decreased, SOD and the GSH/GSSG ratio increased in a dose-dependent manner. In the MaR1-treated AP mice, inflammation of the pancreas and the expression of inflammatory cytokines, pancreatic acinar cell apoptosis, Bcl-2 expression, and expression of TLR4, MyD88, and p-NF-kappaB p65 were reduced, but Bax, cleaved caspase-3, and cleaved caspase-9 expression increased. CONCLUSIONS In a mouse model of caerulein-induced AP, treatment with MaR1 reduced oxidative stress and inflammation and reduced apoptosis.

Multiple pregnancies and the risk of diabetes mellitus in postmenopausal women.

OBJECTIVE: We aimed to investigate whether the number of pregnancies during childbearing age was associated with diabetes in postmenopausal women with no history of gestational diabetes. METHODS: Our data source was the continuous National Health and Nutrition Examination Survey 1999 to 2014. We selected 9,138 postmenopausal women over 40 years old who did not have a history of gestational diabetes during pregnancy. Logistic regression analyses were applied for the association of the number of pregnancies with diabetes. RESULTS: We found women with ≥4 pregnancies had significantly greater fasting plasma glucose (FPG), glycated hemoglobin (HbA1c), 2-hour plasma glucose, and the Homeostatic Model Assessment of Insulin Resistance than those with two to three pregnancies (all P < 0.01). These women also had a significantly higher prevalence of diabetes (28.4% vs 20.7%; P < 0.001). Using the two to three pregnancies group as the reference, we observed a positive association of log-FPG and log-HbA1c with 4 or more pregnancies after adjustment for sociodemographic, lifestyle, and reproductive factors, and body mass index (both P < 0.05). Compared to women with two to three pregnancies, the odds ratios for diabetes were 1.31 (95% confidence interval [CI] 1.01-1.71) for women who never got pregnant and 1.28 (95% CI 1.10-1.48) for those with at least 4 pregnancies after multivariate adjustment. CONCLUSIONS: At least 4 pregnancies through childbearing age may be a potential risk factor for diabetes in postmenopausal women without a history of gestational diabetes.

Clinical characteristics and surgical outcome in children with intussusceptions secondary to pathologic lead points: retrospective study in a single institution.

BACKGROUND: Intussusception secondary to pathologic lead points (PLPs) is a potential surgical emergency and almost all cases need surgery. The aim of this study was to evaluate the clinical manifestations, physical examinations and surgical outcomes of secondary intussusception (SI) caused by PLPs, as well as to improve the diagnosis and treatment of PLPs in children and infants. MATERIALS AND METHODS: We retrospectively reviewed the records of 83 children and infants who were diagnosed with intussusception secondary to PLPs in our institution. The ultimate diagnosis was dependent on histopathological findings under a microscope by a pathologist. Patients were divided into a younger group (< 2 years old) and the older group (> 2 years old) according to age. Patient demographics, clinical manifestations, duration of symptoms, auxiliary examinations, and the presence of pathological lead point were recorded. RESULTS: A total of 83 patients were found with intussusception secondary to PLPs in this study. Patients were aged from 4 days to 14 years, with a mean age of 3.8 years (median 1.5; range 0-14 years). There were 47 cases in the younger group and 36 cases in the older group. The main clinical symptoms were intermittent crying or abdominal pain. PLPs were observed in only ten patients on US (12%). Ten patients underwent enteroscopy examination for further diagnosis, and all the patients had positive findings including seven cases of Peutz-Jeghers syndrome and three cases of benign polyps. Technetium-99 m pertechnetate scans were performed in ten patients and five patients had positive results (50%). Based on the surgical findings, complex/compound is the most common type of intussusception, followed by small intestinal and ileo-colic type. The main types of PLPs were Meckel's diverticulum (n = 31), duplication cyst (n = 19) and benign polyps (n = 13). Meckel's diverticulum and intestinal duplication were the most common causes of secondary intussusception among children younger than 2 years, accounting for 81% (38/47) of the cases. The most common causes of secondary intussusception in children older than 2 years were intestinal polyps, Meckel's diverticulum and Peutz-Jeghers syndrome, accounting for 72% (26/36) of the cases. CONCLUSIONS: The presence of a pathological lead point is more likely in older children. The most common types of intussusception secondary to PLPs are complex/compound and small intestinal. Meckel's diverticulum and intestinal duplication were the most common causes of secondary intussusception among younger children and Peutz-Jeghers syndrome and intestinal polyps were commonly seen in older children.

Enhanced anti-tumor effects of doxorubicin on glioma by entrapping in polybutylcyanoacrylate nanoparticles.

For effective therapy for glioma, it is essential for chemotherapeutics to pass the blood-brain barrier to target glioma cells with little side effects to surrounding normal cells. In this study, we prepared doxorubicin-polybutylcyanoacrylate nanoparticles (Dox-PBCA-NP) and assessed its inhibition effects on glioma both in vitro and in vivo. Dox-PBCA-NP was prepared using the emulsion polymerization method. The size and size distribution of nanoparticles were measured by Malven laser mastersizer and the morphology was observed under transmission electron microscope. Drug loading (DL) and entrapment efficiency (EE) of doxorubicin in the nanoparticles were measured by UV spectra. The proliferation of C6 glioma cells was detected by MTT assay, and cell cycle was analyzed by flow cytometry. The expression of telomerase was detected by immunocytochemical analysis. The anti-tumor efficiency of Dox-PBCA-NP was assessed in C6 glioma intracranial implant rat model. The average diameter of NP-Dox was 120 nm, DL was 10.58 %, and EE was 87.43 %. We found that the cytotoxicity of Dox-PBCA-NP was lower than Dox in vitro. In vivo, Dox-PBCA-NP could transport more Dox into tumors compared to contralateral control, and the life span was longer than Dox. Moreover, Dox-PBCA-NP had less cardiotoxicity than Dox. Taken together, our results suggest that Dox-PBCA-NP exhibits better therapeutic effects against glioma and fewer side effects and is a potential nano-scale drug delivery system for glioma chemotherapy.