Pancreatic cancer is among the most malignant cancers,and thus early intervention is the key to better survival outcomes.However,no methods have been derived that can reliably identify early precursors of development into malignancy.Therefore,it is urgent to discover early molecular changes during pancreatic tumorigenesis.As aberrant glycosylation is closely associated with cancer progression,numerous efforts have been made to mine glycosylation changes as biomarkers for diagnosis;however,detailed glycoproteomic information,especially site-specific N-glycosylation changes in pancreatic cancer with and without drug treatment,needs to be further explored.Herein,we used comprehensive solid-phase chemoenzymatic glycoproteomics to analyze glycans,glycosites,and intact glycopeptides in pancreatic cancer cells and patient sera.The profiling of N-glycans in cancer cells revealed an increase in the secreted glycoproteins from the primary tumor of MIA PaCa-2 cells,whereas human sera,which contain many secreted glycoproteins,had significant changes of glycans at their specific glycosites.These results indicated the potential role for tumor-specific glycosylation as disease biomarkers.We also found that AMG-510,a small molecule inhibitor against Kirsten rat sarcoma viral oncogene homolog(KRAS)G12C mutation,profoundly reduced the glycosylation level in MIA PaCa-2 cells,suggesting that KRAS plays a role in the cellular glycosylation process,and thus glycosylation inhibition contributes to the anti-tumor effect of AMG-510.

Cardiac-resident macrophages (CRMs) play important roles in homeostasis, cardiac function, and remodeling. Although CRMs play critical roles in cardiac regeneration of neonatal mice, their roles are yet to be fully elucidated. Therefore, this study aimed to investigate the dynamic changes of CRMs during cardiac ontogeny and analyze the phenotypic and functional properties of CRMs in the promotion of cardiac regeneration. During mouse cardiac ontogeny, four CRM subsets exist successively: CX₃CR1⁺CCR2^-Ly6C^-MHCII^- (MP1), CX₃CR1^lowCCR2^lowLy6C^-MHCII^- (MP2), CX₃CR1^-CCR2⁺Ly6C⁺MHCII^- (MP3), and CX₃CR1⁺CCR2^-Ly6C^-MHCII⁺ (MP4). MP1 cluster has different derivations (yolk sac, fetal liver, and bone marrow) and multiple functions population. Embryonic and neonatal-derived-MP1 directly promoted cardiomyocyte proliferation through Jagged-1-Notch1 axis and significantly ameliorated cardiac injury following myocardial infarction. MP2/3 subsets could survive throughout adulthood. MP4, the main population in adult mouse hearts, contributed to inflammation. During ontogeny, MP1 can convert into MP4 triggered by changes in the cellular redox state. These findings delineate the evolutionary dynamics of CRMs under physiological conditions and found direct evidence that embryonic and neonatal-derived CRMs regulate cardiomyocyte proliferation. Our findings also shed light on cardiac repair following injury.

Antibodies, Antiphospholipid ; Antiphospholipid Syndrome ; Female ; Humans ; Killer Cells, Natural ; Pregnancy ; Pregnancy Complications

Objective:To explore the mechanism of B10 cell involved in cardiomyocyte hypertrophy following myocarditis, and to develop potential therapeutic strategies.Methods:BALB/c mice infected with Coxsackie virus B3 induced viral myocarditis model. The expression of angiotensin (ANG)Ⅱ and its receptor in myocarditis mice was detected. The changes of B10 cells in the hearts of control mice and myocarditis mice were analyzed by flow cytometry. After losartan was administered to myocarditis mice, the degree of myocardial inflammation was detected by HE staining, the expression of inflammatory factors was detected by ELISA, the myocardial hypertrophy was detected by wheat germ agglutinin (WGA) staining, and the changes of B10 cells in the heart were analyzed by flow cytometry. The levels of cardiac troponin T (C-TNT) and high mobility group box 1 (HMGB1) protein in neonatal mouse cardiomyocytes treated with ANGⅡ and ANGⅡ+ IL-10 were detected. Cardiomyocytes were treated with ANGⅡ, ANGⅡ+ B10 cells, ANGⅡ+ B10 cells + IL-10 receptor antibody and ANGⅡ+ B cells to detect C-TNT protein levels, and Annexin-V/PI was used to detect the apoptosis of cardiomyocytes. Cardiomyocytes were treated with oxidized HMGB1, reduced HMGB1 and disulfide HMGB1, and C-TNT expression was detected.Results:Coxsackievirus B3 infection caused cardiac hypertrophy, high expression of ANGⅡ and its receptor, and transient increase of B10 cells in mice. Losartan treatment blocked the angiotensin receptor, reduced expansion of B10 cells. B10 cells alleviated apoptosis of cardiomyocytes and inhibited the production of HMGB1 induced by ANGⅡ patch by producing IL-10, thus alleviating viral myocarditis and cardiac hypertrophy.Conclusions:B10 cells may play an important role in myocardial protection in myocarditis.

Objective:To investigate and analyze the mutation of homeobox D13 (HOXD13) gene in a pedigree with synpolydactyly.Methods:The data of a pedigree with synpolydactyly who were treated by the First Affiliated Hospital of Fujian Medical University in August 2013 were collected. The proband and all members were diagnosed based on clinical manifestations, imaging, and family history. Genomic DNA was extracted from peripheral blood of some members in the pedigree. The HOXD13 gene was identified by polymerase chain reaction (PCR) and DNA sequence analysis.Results:Among the 35 members of this 4-generation family, 12 patients were affected(5 males and 7 females), and the proband was a 20 years old female patient. All the patients had bilateral 3rd and 4th syndactyly with clinodactyly, which could not be extended, and the remaining fingers could move freely. The foot showed unilateral or bilateral 4th and 5th syndactyly with polydactyly in soft tissue web, which was consistent with the typical autosomal dominant synpolydactyly phenotypic characteristics. The analysis of HOXD13 gene in 19 family members (12 normal members and 7 patients) showed that the nucleotide sequences of HOXD13 gene in 12 normal members were normal, and 9 alanine residues were inserted into the polyalanine chain of the first exon 1 of HOXD13 gene in the 7 patients, which extended the alanine residue base from the normal 15 to 24.Conclusions:The pathogenic mutation of HOXD13 gene in this pedigree was the extension mutation of polyalanine chain, which resulted in the typical synpolydactyly.

Objective:To investigate and analyze the mutation of homeobox D13 (HOXD13) gene in a pedigree with synpolydactyly.Methods:The data of a pedigree with synpolydactyly who were treated by the First Affiliated Hospital of Fujian Medical University in August 2013 were collected. The proband and all members were diagnosed based on clinical manifestations, imaging, and family history. Genomic DNA was extracted from peripheral blood of some members in the pedigree. The HOXD13 gene was identified by polymerase chain reaction (PCR) and DNA sequence analysis.Results:Among the 35 members of this 4-generation family, 12 patients were affected(5 males and 7 females), and the proband was a 20 years old female patient. All the patients had bilateral 3rd and 4th syndactyly with clinodactyly, which could not be extended, and the remaining fingers could move freely. The foot showed unilateral or bilateral 4th and 5th syndactyly with polydactyly in soft tissue web, which was consistent with the typical autosomal dominant synpolydactyly phenotypic characteristics. The analysis of HOXD13 gene in 19 family members (12 normal members and 7 patients) showed that the nucleotide sequences of HOXD13 gene in 12 normal members were normal, and 9 alanine residues were inserted into the polyalanine chain of the first exon 1 of HOXD13 gene in the 7 patients, which extended the alanine residue base from the normal 15 to 24.Conclusions:The pathogenic mutation of HOXD13 gene in this pedigree was the extension mutation of polyalanine chain, which resulted in the typical synpolydactyly.

Objective: To explore the clinical application of subpectoral silicone implant associated with autologous fat grafting after tissue expansion, for breast reconstructions. Methods: From Jan 2013 to Dec 2016, a total of 15 female patients were admitted to the plastic surgery department of the First Affiliated hospital of Fujian Medical University. They were after or were prepared to unilateral modified radical mastectomy. Patients were aged 18-50 years old, with the average of 33.5 years. There were 8 cases of T₁N₀M_O (T_1micN₀M_O, n=1), 3 cases of T₂N₀M_O, 1 case of T₂N₁M_O, 1 case of T₁N₂M_O, 1 case of T₂N₂M_O and 1 case of T₃N₃M_O. A tissue expander was inserted beneath the posterior pectoralis major in first surgery. The cavity stripped range is greater than the expander around 1cm. The lowest position of the expander was determined according to the IMF of the other breast. The tissue expander was gradually filled intraoperatively and postoperatively periodically, until the expander exceeds 30%-50% of the volume of the other breast. Three months later, as the soft tissue over the breast area has stretched enough, the expander was removed in a second operation and a permanent implant was inserted. Meanwhile, the autologous fat particles, harvested from thigh or abdomen, was centrifuged at 800×g for 3 minutes. Then it was injected into the subcutaneous and posterior pectoralis major space. And the other breast may be adjusted if necessary for symmetry. Results: Fifteen patients was included in this study (immediately tissue expander implanted, n=8; delayed tissue expander implanted, n=7). All the expanders are round in shape. The interval between the two operations was 6-12 months, with an average of 7.4 months. The total salt-water solution injected was 310-450 ml per patient, with an average of 365 ml. The volume of breast implants was 180-280 ml. The injection volume of autologous fat was 65-230 ml, 122.7 ml on average. All incisions healed well. The follow-up period was 1-3 years, with an average of 2 years. The reconstructed breasts were in a natural shape, with a slight drop and bilateral symmetry. According to Harris breast shape evaluation criteria, all reconstructed breasts were graded as excellent. Conclusions The permanent implant associated with autologous fat grafting after tissue expansion is alternative for breast reconstruction.

Objective The aims of this study were to construct short hairpin RNA(shRNA)lentiviral vector in breast cancer T47D cells,to carry out RNA interference on lysine acetyltransferase 6B(KAT6B/MORF)gene,to down-regulate its expression and to explore its function.Methods Two pairs of single-stranded short hairpin RNA(shRNA5 and shRNA8)and the corresponding control sequences(Scramble5 and Scramble8)were synthesized based on the CDS of KAT6B gene.Polymerase chain reaction(PCR) was used to amplify double-stranded and ligated with the entry vector(pENTR/pSM2(CMV)GFP),which were subjected to a doub-le digestion(EcoRl and Xhol)linearization and homologous recombination with the entry vector(pENTR/pSM2(CMV)GFP)to obtain an entry clone containing the desired fragment.The target fragment was recombined onto the target vector(pLenti x1 puro DEST)via the LR cloning reaction of the Gateway system.The lentiviral packaging plasmids were co-transfected into HEK-293T cells with two pairs of target plasmids. The supernatant of HEK -293T cells was collected and transformed into T47D cells. The expression of KAT6B protein was detected in T47D cells by Western blot.Results The single colony obtained from the transformation was identi-fied by sequencing,which was consistent with the target sequence,indicating that the lentiviral vector had been successfully construc-ted.The expression of KAT6B protein was significantly lower in the shRNA KAT6B group than that in the control group,which indica-ted that the constructed gene silencing vector could play a role in the KAT6B gene in T47D cells.Conclusion The shRNA lentiviral gene silencing vectors of KAT6B were constructed and identified in T47D cells,which indicated that the foundation for further study

Objective To investigate the role of miR-592 in the Glioma.Methods We first analyzed the expression of miR-592 in Glioma tissues from patients by quantitative PCR.We transfected U251 cells with miR-592 mimics and then detected the growth of cells by MTT assay.We performed dual-luciferase reporter assay and western blot assay to examine whether Runx2 was the direct target of miR-592 in U251 cells.In order to test whether Runx2 was the functional target of miR-592,we determined the cell growth curve by down-regulating the level of Runx2.Moreover,we also detected the apoptosis of U251 after Runx2 knockdown.Results The expression of miR-592 was significantly reduced in glioma tissues (t=2.752,P--0.013).Over-expression miR-592 remarkably increased the apoptotic rate of U251 cells compared with the control group (t=2.127,P=0.031;t=2.284,P=0.026).Flow cytometry analysis showed that MiR-592 significantly promoted apoptotic cell death of U251 cells Apoptosis rate was 7.2％±0.68％ in miR-592 group and 17.47％±1.45％ in control group (t=3.294,P=0.007).The results of double luciferase assay and Western blot assay showed that miR-592 directly targeted the 3'Runx2 of-UTR to inhibit the level of Runx2 protein.The effect of down-regulation of Runx2 on the growth of U251 cells was detected,the results showed that growth was significantly slower in the cells transfected with Runx2 siRNA than in those without Runx2 siRNA (t=3.124,P=0.O11).Detection of cycle by flow cytometry showed that runx2 down-regulated the apoptosis rate of U251 cells.Tumor growth curve showed that overexpression of miR-592 significantly inhibited tumor growth and the down regulation of Runx2 expresssion also significantly inhibited tumor growth.Conclusion miR-592 suppresses the growth and promotes the apoptotic rate of U251 cells by targeting Runx2.

Objective To investigate the prognostic value of platelet to lymphocyte ratio (PLR) for early virological response in Entecavir (ETV)-treated chronic hepatitis B (CHB) patients with genotype C infection.Methods Ninety-one genotype C CHB patients with HBV DNA≥1×105 copies/mL were treated with ETV (0.5 mg/d) for 10-13 days.The correlation between PLR and viral load decline was evaluated by Pearson or Spearman's rank correlation coefficient.Stepwise linear regression analysis was used to establish the prediction model of virological response.Receiver operating characteristic (ROC) curves were used to evaluate the predictive value of PLR for early virological response in ETV-treated patients with genotype C hepatitis B virus (HBV) infection.Results After 10-13 days of ETV treatment, HBV DNA decreased ≥1×lg copies/mL from baseline in 89 cases of the 91 patients, while HBV DNA declined ≥2×lg copies/mL in 65 patients and 4 patients achieved HBV DNA<500 copies/mL.HBV DNA decline was positively correlated with baseline PLR levels (r=0.235 09, P<0.05).After adjustment for age, gender, Hepatitis B e Antigen (HBeAg), and treatment days, HBV DNA decline was still positively correlated with baseline PLR levels (r=0.220 26, P<0.05).Area under curve (AUC) of prediction model including age , baseline aspartate transaminase (AST) and HBV DNA was 0.759 (95% CI : 0.660-0.859, P<0.01).After adding PLR to the prediction model, the AUC was 0.780 (95% CI: 0.685-0.875, P<0.01).Conclusions PLR is predictive to early virological response in ETV-treated CHB patients with genotype C infection.Higher baseline PLR level indicates a better virological response.PLR monitoring should be recommended in CHB patients with antiviral treatment in clinical practice.