Objective:The aim of the study was to evaluate the effect of Sr2+doped micro arc oxidation treated surface on osteogen-ic differentiation of BMSCs and osseointegration in the femurs of type 2 diabetes mellitus rats.Methods:TC4 surface was treated by micro arc oxidation to create TiO2 coating,then Sr was incorporated to the coating by electrochemical treated in a strontium dichlo-ride solution.The surface topography was characterized using scanning electronic microscope.Chemical compositions were deter-mined by X-ray photoelectron spectroscopy.Type 2 diabetic rat was induced by feeding with high-fat diet and following injection of streptozotocin.BMSCs were isolated after diabetic rat was euthanized.To study the cells attachment to different samples,cells actin filaments were observed to evaluate cells attachment to different plates.MTT assay was used to evaluate the cell proliferation activity on the plates.Cells differentiation was evaluated by RT-PCR,ALP activity and immunofluorescence of osteocalcin(OCN)protein.MAO and MAO+Sr implants were inserted in the diabetic rats' femurs to evaluate the influence on new bone formation and bone im-plant contact.A software was used to compare the differences of cells proliferation and osteogenic differentiation on different plates,as well as bone regeneration and bone implant contact between the implants with different surfaces.Results:After electrochemical treatment,Sr was detected on the MAO surface.The osteogeneic-related markers were upregulated on the MAO+Sr surface.In vivo study revealed the enhanced bone generation and implant bone contact of MAO+Sr surface in diabetic rats.Conclusion:Compared to the MAO surface,Sr doped micro-arc oxidation treated surface could further improve cells osteogenic differentiation and osseointe gration in diabetics type 2 diabetic rats.

OBJECTIVES: To investigate the role of the calcium/calmodulin (CaM)-mediated activation of calcineurin (CN)-nuclear factor of activated T cells (NFAT) signaling pathway in mediating the regulatory effect of hyperforin (HY) on stress-induced depression-like disorder (DP) in mice. METHODS: C57BL/6J mice were randomly divided into control group, DP model group, and hyperforin treatment group (n=15). Behavioral changes of the mice were assessed using open field test (OFT), sucrose preference test (SPT), tail suspension test (TST), light/dark box test (LDB), and novel object suppression test (NSFT). Immunohistochemistry was used to detect tyrosine hydroxylase (TH) expression in the CA1 region of the hippocampus, and serum serotonin (5-HT) and norepinephrine (NA) levels were detected with ELISA. Western blotting was used to analyze the expressions of TNF-α, IL-1β, IL-2, and CN-NFAT pathway proteins. In cultured BV-2 microglial cells with lipopolysaccharide (LPS) stimulation, the effects of hyperforin and CN inhibitor (CNIS) on expressions of ionized calcium-binding adapter molecule 1 (IBA-1), 5-HT, NA, inflammatory cytokines and CN-NFAT pathway proteins were examined using immunofluorescence assay, ELISA or Western blotting. RESULTS: Compared with the control mice, the mice in DP group showed significantly reduced activity in OFT, decreased sucrose consumption in SPT, reduced shuttle crossing in LDB, and lowered food intake in NSFT with significantly increased immobility in TST. The mice with DP showed significantly decreased TH-positive neurons, lowered 5-HT and NA levels, and increased expressions of TNF-α, IL-1β, IL-2 and CaM-CN-NFAT pathway proteins. In cultured BV-2 cells, LPS stimulation strongly increased cellular IBA-1 expression, decreased the levels of neurotransmitters (5-HT and NA), and increased the levels of inflammatory cytokines and CN-NFAT signaling, and these changes were effectively reversed by treatment with hyperforin or CNIS. CONCLUSIONS Hyperforin improves stress-induced depression-like behaviors in mice and activated BV-2 cells by targeting the CN-NFAT signaling pathway.
Animals ; Mice, Inbred C57BL ; Mice ; Microglia/drug effects* ; Depression/etiology* ; Perylene/pharmacology* ; Calcineurin/metabolism* ; NFATC Transcription Factors/metabolism* ; Calcium Signaling/drug effects* ; Stress, Psychological ; Phloroglucinol/pharmacology* ; Signal Transduction ; Male ; Behavior, Animal/drug effects* ; Terpenes

Objective:The aim of the study was to evaluate the effect of Sr2+doped micro arc oxidation treated surface on osteogen-ic differentiation of BMSCs and osseointegration in the femurs of type 2 diabetes mellitus rats.Methods:TC4 surface was treated by micro arc oxidation to create TiO2 coating,then Sr was incorporated to the coating by electrochemical treated in a strontium dichlo-ride solution.The surface topography was characterized using scanning electronic microscope.Chemical compositions were deter-mined by X-ray photoelectron spectroscopy.Type 2 diabetic rat was induced by feeding with high-fat diet and following injection of streptozotocin.BMSCs were isolated after diabetic rat was euthanized.To study the cells attachment to different samples,cells actin filaments were observed to evaluate cells attachment to different plates.MTT assay was used to evaluate the cell proliferation activity on the plates.Cells differentiation was evaluated by RT-PCR,ALP activity and immunofluorescence of osteocalcin(OCN)protein.MAO and MAO+Sr implants were inserted in the diabetic rats' femurs to evaluate the influence on new bone formation and bone im-plant contact.A software was used to compare the differences of cells proliferation and osteogenic differentiation on different plates,as well as bone regeneration and bone implant contact between the implants with different surfaces.Results:After electrochemical treatment,Sr was detected on the MAO surface.The osteogeneic-related markers were upregulated on the MAO+Sr surface.In vivo study revealed the enhanced bone generation and implant bone contact of MAO+Sr surface in diabetic rats.Conclusion:Compared to the MAO surface,Sr doped micro-arc oxidation treated surface could further improve cells osteogenic differentiation and osseointe gration in diabetics type 2 diabetic rats.

Objective:To construct an inpatient-outpatient-home step swallowing rehabilitation training program for partial laryngectomy patients.Methods:From June 2022 to May 2023, a preliminary program for inpatient-outpatient-home step swallowing rehabilitation training for partial laryngectomy patients was developed through literature search and analysis. Delphi method was used for two rounds of expert inquiry on the inpatient-outpatient-home step swallowing rehabilitation training program for partial laryngectomy patients.Results:A total of 15 experts were included for two rounds of expert inquiry. The effective response rates of the questionnaires from the two rounds of expert inquiries were 100.0% (15/15). The expert authority coefficients for the two rounds of inquiry were 0.885 and 0.855, respectively, and the Kendall harmony coefficients were 0.217 and 0.230, respectively ( P<0.01). The final inpatient-outpatient-home step swallowing rehabilitation training program for partial laryngectomy patients included three primary items (swallowing training step, swallowing training methods, and feeding guidance methods), 21 secondary items, and 35 tertiary items. Conclusions:The inpatient-outpatient-home step swallowing rehabilitation training program for partial laryngectomy patients based on the Delphi method has certain scientific and feasibility and can provide a reference for medical and nursing staff.

Objective To investigate the effect and mechanism of Congrong Shujing Granules on promoting microglial polarization and inhibiting neuroinflammation through the nucleotide-binding oligomeric domain-like receptor protein 3(NLRP3)/Caspase-1 signaling pathway.Methods Twenty Sprague-Dawley rats were assigned to the blank serum and Congrong Shujing Granules containing serum groups using random number table method,with 10 rats in each group.Rats in the Congrong Shujing Granules containing serum group received intragastric administration of Congrong Shujing Granules(2.57 g/kg)and the rats in the blank serum group received intragastric administration of physiological saline of equal volume.Blank serum and Congrong Shujing Granules containing serum were prepared separately.Mouse microglia cells BV-2 were cultured in vitro,and the optimal concentration of 1-methyl-4-phenylpyridine(MPP+)and optimal volume fraction of Congrong Shujing Granules containing serum were selected by the CCK-8 assay and immunofluorescence staining.And the NLRP3 inhibitor MCC950 was used as a postive control.Cells were divided into the blank serum group(10%blank serum),model group(10%blank serum+500 μmol/L MPP+),Congrong Shujing Granules containing serum group(10%Congrong Shujing Granules containing serum+500 μmol/L MPP+),and MCC950 group(10%blank serum+10 μmol/L MCC950+500 μmol/L MPP+),and intervened separately.After 14 h of intervention,morphological changes in BV-2 cells were observed.The contents of interleukin(IL)-1β,tumor necrosis factor-α(TNF-α),IL-6,and IL-4 were detected by an enzyme-linked immunosorbent assay.The mRNA expressions of differentiation cluster 86(CD86),inducible nitric oxide synthase(iNOS),CD206,and arginase 1(Arg1)were detected by real-time fluorescence PCR.The expressions of CD86,Arg1,Ionized calcium binding adaptor molecule 1(Iba1),and NLRP3 were detected by immunofluorescence staining.The expression of iNOS,Arg1,TNF-α,IL-6,IL-4,NLRP3,pro-cysteinyl aspartate specific proteinase 1(pro-Caspase-1),and Caspase-1 proteins was detected by Western blotting.Results Iba1 activation and expression increased under the MPP+(12 h,500 μmol/L)intervention(P<0.05),and cell viability was not affected.There was no statistically significant effect on cell viability after treatment with 10%Congrong Shujing Granules containing serum alone or in combination with MPP+(P>0.05).Compared to the blank serum group,BV-2 cells in the model group showed multiple branches and protruded in the shape of an amoeba.The contents of IL-1β,TNF-α,and IL-6 increased,while the contents of IL-4 decreased.The mRNA expressions of CD86 and iNOS increased,while mRNA expressions of CD206 and Arg1 decreased.The mean fluorescence intensity of CD86,Iba1,and NLRP3 increased,while the mean fluorescence intensity of Arg1 decreased.The protein expressions of iNOS,TNF-α,IL-6,NLRP3,pro-Caspase-1,and Caspase-1 increased,while the protein expressions of Arg1,IL-4 decreased,P<0.05.Compared to the model group,the Congrong Shujing Granules containing serum group and MCC950 group showed a decrease in the branch of cell protrusions,reduced cell activation,decreased levels of IL-1β,TNF-α,and IL-6,increased levels of IL-4,decreased expression of CD86 and iNOS mRNA,increased expression of CD206 mRNA,the decreased mean fluorescence intensity of CD86,Iba1,and NLRP3,the increased mean fluorescence intensity of Arg1,decreased expression of iNOS,TNF-α,IL-6,NLRP3,pro-Caspase-1,and Caspase-1 proteins,and increased expression of Arg1 and IL-4 proteins,P<0.05.Conclusion Congrong Shujing Granules containing serum may alleviate the MPP+-induced neuroinflammatory response by inhibiting the NLRP3/Caspase-1 signaling pathway to regulate M1/M2 phenotype polarization of microglia.

Objective To investigate the effect and mechanism of Congrong Shujing Granules on promoting microglial polarization and inhibiting neuroinflammation through the nucleotide-binding oligomeric domain-like receptor protein 3(NLRP3)/Caspase-1 signaling pathway.Methods Twenty Sprague-Dawley rats were assigned to the blank serum and Congrong Shujing Granules containing serum groups using random number table method,with 10 rats in each group.Rats in the Congrong Shujing Granules containing serum group received intragastric administration of Congrong Shujing Granules(2.57 g/kg)and the rats in the blank serum group received intragastric administration of physiological saline of equal volume.Blank serum and Congrong Shujing Granules containing serum were prepared separately.Mouse microglia cells BV-2 were cultured in vitro,and the optimal concentration of 1-methyl-4-phenylpyridine(MPP+)and optimal volume fraction of Congrong Shujing Granules containing serum were selected by the CCK-8 assay and immunofluorescence staining.And the NLRP3 inhibitor MCC950 was used as a postive control.Cells were divided into the blank serum group(10%blank serum),model group(10%blank serum+500 μmol/L MPP+),Congrong Shujing Granules containing serum group(10%Congrong Shujing Granules containing serum+500 μmol/L MPP+),and MCC950 group(10%blank serum+10 μmol/L MCC950+500 μmol/L MPP+),and intervened separately.After 14 h of intervention,morphological changes in BV-2 cells were observed.The contents of interleukin(IL)-1β,tumor necrosis factor-α(TNF-α),IL-6,and IL-4 were detected by an enzyme-linked immunosorbent assay.The mRNA expressions of differentiation cluster 86(CD86),inducible nitric oxide synthase(iNOS),CD206,and arginase 1(Arg1)were detected by real-time fluorescence PCR.The expressions of CD86,Arg1,Ionized calcium binding adaptor molecule 1(Iba1),and NLRP3 were detected by immunofluorescence staining.The expression of iNOS,Arg1,TNF-α,IL-6,IL-4,NLRP3,pro-cysteinyl aspartate specific proteinase 1(pro-Caspase-1),and Caspase-1 proteins was detected by Western blotting.Results Iba1 activation and expression increased under the MPP+(12 h,500 μmol/L)intervention(P<0.05),and cell viability was not affected.There was no statistically significant effect on cell viability after treatment with 10%Congrong Shujing Granules containing serum alone or in combination with MPP+(P>0.05).Compared to the blank serum group,BV-2 cells in the model group showed multiple branches and protruded in the shape of an amoeba.The contents of IL-1β,TNF-α,and IL-6 increased,while the contents of IL-4 decreased.The mRNA expressions of CD86 and iNOS increased,while mRNA expressions of CD206 and Arg1 decreased.The mean fluorescence intensity of CD86,Iba1,and NLRP3 increased,while the mean fluorescence intensity of Arg1 decreased.The protein expressions of iNOS,TNF-α,IL-6,NLRP3,pro-Caspase-1,and Caspase-1 increased,while the protein expressions of Arg1,IL-4 decreased,P<0.05.Compared to the model group,the Congrong Shujing Granules containing serum group and MCC950 group showed a decrease in the branch of cell protrusions,reduced cell activation,decreased levels of IL-1β,TNF-α,and IL-6,increased levels of IL-4,decreased expression of CD86 and iNOS mRNA,increased expression of CD206 mRNA,the decreased mean fluorescence intensity of CD86,Iba1,and NLRP3,the increased mean fluorescence intensity of Arg1,decreased expression of iNOS,TNF-α,IL-6,NLRP3,pro-Caspase-1,and Caspase-1 proteins,and increased expression of Arg1 and IL-4 proteins,P<0.05.Conclusion Congrong Shujing Granules containing serum may alleviate the MPP+-induced neuroinflammatory response by inhibiting the NLRP3/Caspase-1 signaling pathway to regulate M1/M2 phenotype polarization of microglia.

Maternal diabetes not only affects the mother′s own health but also has a significant influence on offspring.Adverse prenatal and intrauterine condition can influence fetal kidney development, which may increase the risk of kidney disease in the adulthood.This article reveiws the impact of maternal diabetes on kidney structure and function.Maternal diabetes can increase the risk of congenital anomalies of the kidney and urinary tract(CAKUT)and impair early and long-term renal function of offspring.In addition, this article reveiws the research progress of the potential mechanisms of how maternal diabetes affects kidney development including oxidative stress, key signal pathway changes of kidney development and epigenetic changes.

Objective:To study factors associated with postoperative acute pancreatitis (POAP) in patients following pancreaticoduodenectomy.Methods:This retrospective analysis included 60 patients who underwent pancreaticoduodenectomy at the First Affiliated Hospital of Soochow University from Jan 2020 to Aug 2021. Enhanced computed tomography was used to identify POAP during postoperative period of 4 to 9 days. Univariate analysis and multivariate analysis were used to find out the risk factors of POAP.Results:Of the 60 patients, 13 cases (21.7%) developed POAP. The incidence of clinically related pancreatic fistula with abdominal abscess (76.9% vs.19.1%, χ2=15.71, P<0.000 1), postoperative hospital stay (26 d vs. 18 d, U=141.5, P=0.002 5) and the severity of complications (Clavien-Dindo grade≥Ⅲ: 53.8% vs. 21.3%, χ2=5.32, P=0.02) were significantly higher in the POAP group. But there was no significant deviation between the two groups when it comes to the severe post pancreatectomy delayed hemorrhage (7.7% vs. 0, χ2=3.68, P=0.06) and the delayed gastric emptying (30.8% vs. 21.3%, χ2=0.51, P=0.47). In the univariate analysis, patients with higher body mass index ( P=0.000 3), smaller main pancreatic duct diameter ( P<0.000 1) and softer texture of the pancreas ( P=0.009) were more likely to develop POAP after pancreaticoduodenectomy. In the multivariate analysis, the pancreatic duct diameter≤2 mm ( OR=0.005,95% CI 0.000 06-0.44, P=0.020), the softer texture of pancreas ( OR=0.005, 95% CI 0.000 04-0.47, P=0.023) were risk factors for POAP. Conclusions:Patients with postoperative acute pancreatitis increased the incidence of pancreatic fistula complicating abdominal abscess.Small caliber pancreatic tube, soft texture of pancreas were risk factors of POAP.

Objective:To investigate the clinical characteristics and treatment of the abdominal cocoon.Mehods:The clinical data of 28 patients with abdominal cocoon from Jan 2004 to Dec 2018 were analyzed retrospectively.Results:Intestinal obstruction was the main clinical manifestations (25 cases), recurrent chronic ileus(17 cases) and abdominal mass (7 cases). Preoperative imaging examination showed varying degrees of intestinal obstruction. CT or MRI scan displayed that small intestinal loops were disorganized , clustered and encased in a thickened capsule. All the cases underwent operations, showing that small bowel were encapsulated in a dense gray-white fibrous membrane. Adhesiolysis and fibrous membrane excision were done with segmental enterectomy when it was necessary. Early postoperative intestinal obstruction occured in 6 cases, all were cured by conservative treatment.Conclusions:The combination of clinical symptoms and CT or MRI may facilitate in preoperative diagnosis. Abdominal cocoon is putative diagnosis when recurrent intestinal obstruction with abdominal mass. Surgery is the therapy of choice.

Objective: To investigate the effect of long-chain non-coding RNA Fez family zinc finger protein 1 antisense RNA1 (lncRNA FEZF1-AS1) on the biological function of hepatocellular carcinoma (HCC). Methods: SMMC771 and BEL-7402 cells were transfected with sh-FEZF1-AS1 and OE-FEZF1-AS1, respectively. The expression of lncRNA FEZF1-AS1 was detected by real-time quantitative PCR. Cell proliferation was detected by Cell Counting Kit-8 (CCK-8), and apoptosis was detected by flow cytometry. The effects of lncRNA FEZF1-AS1 on invasion and migration were detected by Transwell and wound healing assays. The expression levels of adhesion molecules were detected by Western blot. The effect of lncRNA FEZF1-AS1 on the in vivo growth was verified by nude mice xenograft experiments. Results: The silencing or ectopic expression of lncRNA FEZF1-AS1 inhibited or promoted the proliferation of hepatocellular carcinoma cells. CCK-8 assay showed that the proliferation abilities of SMMC7721 and BEL-7402 cells in sh-FEZF1-AS1 transfection group significantly decreased, achieving (35.43±4.06)% and (34.68±3.97)%, respectively, on the fifth day. There were significant differences between sh-FEZF1-AS1 group and sh-NC group [52.21±8.46)% and (53.76±7.64)%] (all P<0.05). In contrast, the proliferation ability of SMMC7721 and BEL-7402 cells transfected with OE-FEZF1-AS1 was significantly increased, achieving (83.49±6.92)% and (80.31±3.13)%, respectively, on the fifth day. There were significant differences between OE-FEZF1-AS1 and OE-NC group [53.03±8.84)% and (55.11±7.09)%] (all P<0.05). The subsequent flow cytometry results showed that cell apoptotic rates of SMMC7721 and BEL-7402 cells transfected with sh-FEZF1-AS1 were (13.02±1.38)% and (11.88±1.29)%, respectively, which were significantly higher than those in sh-NC groups [(5.57±1.46)% and (8.06±1.42)%, respectively, all P<0.05]. In contrast, the apoptotic rates of SMMC7721 and BEL-7402 cells transfected with OE-FEZF1-AS1 were (3.01±0.39)% and (3.22±0.43)%, which were significantly lower than those in OE-NC groups [(6.68±0.96)% and (6.63±0.45)%, all P<0.05]. In addition, knockdown or overexpression of lncRNA FEZF1-AS1 expression inhibited or enhanced the migration and invasion abilities as well as the levels of adhesion molecules in hepatocellular carcinoma cells. After 30 days of feeding under the same conditions, the tumor volumes of sh-FEZF1-AS1 and sh-NC SMMC7721 cells xenograft mice models were (0.26±0.03) cm³ and (0.63±0.06) cm³, respectively, showing significant difference (P<0.05). The tumor volumes of sh-FEZF1-AS1 and sh-NC BEL-7402 cells were (0.31±0.02) cm³ and (0.72±0.08) cm³, and the difference was statistically significant (P<0.05). Conclusion lncRNA FEZF1-AS1 may strengthen the growth, migration and invasion of hepatocellular carcinoma cells.