Neuroendocrine tumors of the gallbladder(GB-NET) are rare, and it lacks early clinical manifestations and has no specific tumor markers, it is difficult to distinguish GB-NET from gallbladder adenocarcinoma. The diagnosis of GB-NET is based on histopathology of the tumor and the assessment of proliferation fraction, which makes it difficult to achieve early diagnosis. GB-NET has a high degree of malignancy, 32.39% of patients have liver metastases at diagnosis, and 51.60% of patients have lymph node metastases, the median survival time is 9 to 10 months.There are currently no specific guidelines or consensus for the treatment of GB-NET. The treatment strategies are choosen mainly by the principles of gallbladder adenocarcinoma. We reviews the clinical and basic researches of GB-NET and case reports from China and across the world, as well as the data from SEER database, and we discuss the research progress on the classification, clinicopathological features, diagnosis, treatment advances and the prognosis.

Objective: The study was designed to evaluate the changes and significance of circulating CD4~+CD25~+ and CD8~+CD28~- regulatory T cells (Tregs) in patients with multiple myeloma (MM).Methods:CD4~+CD25~+ and CD8~+CD28~-Tregs in peripheral blood of 38 patients with MM and of 20 healthy doners were measured by flow cytometry.Serum albumin and β_2-MG in patients with MM were measured using bromocresol green method,transmission turbidimetry respectively.Results:Compared to those of the controls,the proportions of CD4~+CD25~(+/high),CD4~+CD25~(high) CD127~(low) and CD8~+CD28~-Treg cells in newly diagnosed MM patients were elevated.Furthermore,the proportions of CD4~+CD25~(high) and CD4~+CD25~(high)CD127~(low) Tregs in each clinical stage were elevated when compared to those of the controls.The number of the Tregs were increasing with clinical stages and were significantly higher in stage Ⅲ MM than in stageⅠ MM;In stageⅡand Ⅲ MM,there were also elevated proportions of CD8~+CD28~- Tregs,increasing with clinical stages.However,there were no differences when compared between stage Ⅰ MM and the controls;Both the proportions of CD4~+CD25~(+/high) and CD4~+CD25~(high)CD127~(low) Tregs in active MM were not different from stable MM,although all of them were higher than those of controls.The proportion of CD8~+CD28~- Tregs was higher in active MM than in stable MM and controls,but there were no differences when compared between active and stable MM.The proportions of both CD4~+CD25~(high) Tregs and CD4~+CD25~(high)CD127~(low)Tregs had negative correlation with the levels of serum albumin.Conclusion:MM patients have elevated levels of circulating CD4~+CD25~+ and CD8~+CD28~-Tregs,which may be an important mechanism of MM immune evasion,and may be associated with clinical stages,disease progression and prognosis of MM to some extent.

Objective To investigate the characteristics of activation-induced cytidine deaminase (AID) expression level in de novo acute leukemia (AL) patients, chronic myeloid leukemia chronic phase (CML-CP), chronic myeloid leukemia blastic crisis (CML-BC) patients and leukemia cell lines. Methods The expression level of AID mRNA was measured in 89 cases of newly-diagnosed acute lymphoblastic leukemia (ALL) patients, 79 cases of de novo acute myeloid leukemia (AML) patients, 5 cases of CML-BC patients, 5 cases of CML-CP patients and leukemia cell lines NB4, THP-1, KG-1, Raji, K562 by real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR), bone marrow mononuclear cells of 16 normal healthy donors were used as the control group. Results The expression levels of AID mRNA in 89 cases of ALL and 79 cases of AML were 0.006-7 463.175 and 0.005-69.107, the median expression levels were 3.785 and 1.812, the expression level of AID mRNA in the normal control group was 0.146-4.707, and the median expression level was 1.483, respectively. The AID expression levels of ALL, B-ALL, Burkitt leukemia, M4 patients and Raji cells were significantly higher than those of the normal control group (all P <0.05). Nevertheless, the AID mRNA expression levels of M3 patients and NB4, KG-1 cells were lower than those of the normal control group (all P <0.05). Furthermore, the AID mRNA expression levels of K562 cell were strikingly higher than that of the CML-CP patients (P<0.001), so were those of CML-BC, chronic myeloid leukemia myeloid blast crisis (CML-MBC), chronic myeloid leukemia lymphoblastic blast crisis (CML-LBC) patients. Conclusion AID gene shows high expression level in B-ALL, Burkitt leukemia and M4, low expression level in M3 and KG-1 cells, and obvious high expression level in CML-BC.

OBJECTIVETo detect the inflammatory factors of induced sputum (IS) and whole lung lavage fluid in pneumonoconiosis patients and to explore the correlation between the inflammatory factors with pulmonary function.

METHODSThe records of 45 cases of pneumonoconiosis patients were observed. All patients underwent lung function examination, sputum induction and massive whole lung lavage (WLL) sequentially through advance. IS and whole lung lavage fluid were collected respectively. Inflammatory factors of the two specimens were detected by using enzyme linked immunosorbent assay. The correlation of inflammatory factors between the two specimens was analyzed. The relationship between the inflammatory factor and lung function index was observed. The statistical analysis is performed with SPSS 17.0 for Windows. P < 0.05 is considered to be statistically significant.

RESULTSCytokines (MCP-1, TNF-α MIP-1α, NO(2)(-)/NO(3)(-) and IL-16) were significantly associated between IS and whole lung lavage fluid (P < 0.05), while TNF-α, MCP-1, NO(2)(-)/NO(3)(-) and IL-16 were no significantly associated with lung function index (P > 0.05). MIP-1α was significantly associated with FEV(1.0)/VCmax and MEF(25), respectively (P < 0.05).

CONCLUSIONSInflammatory factors were significantly associated between IS and whole lung lavage fluid, which could indicate early lung injury in pneumonoconiosis patients.

Bronchoalveolar Lavage Fluid ; chemistry ; Chemokine CCL2 ; Chemokine CCL3 ; Cytokines ; analysis ; Humans ; Interleukin-16 ; Lung ; Respiratory Function Tests ; Silicosis ; Sputum ; chemistry ; Tumor Necrosis Factor-alpha

Objective:To evaluate effect of diammonium glycyrrhizinate(DG)on lipopolysaccharide(LPS)-induced TLR4/NF-κB inflammatory signaling pathway in BV2 microglial cells and to explore its potential regulatory mechanisms on ameliorating neu-roinflammation.Methods:BV2 microglia injury model was induced by LPS and treated with 20 and 60 μmol/L DG.MTS was used to determine vitality of cells.NO level secreted by cells was detected by Griess.Inflammatory factors TNF-α,IL-6,and IL-1β levels in cell supernatant were measured by ELISA.TNF-α,IL-6,IL-1β,TLR4 and MyD88 mRNA levels were detected by RT-qPCR.Western blot was used to determine expressions of TLR4,MyD88,NF-κB,p-NF-κB,IκB-α and p-IκB-α proteins of cells.Results:Compared with control group,LPS-treated BV2 microglia had significantly lower viability(P<0.05),significantly higher NO secretion(P<0.05),significantly up-regulation levels of inflammatory factors TNF-α,IL-6 and IL-1β(P<0.05),and significantly higher mRNA levels of TNF-α,IL-6,IL-1β,TLR4 and MyD88(P<0.05),TLR4,MyD88,NF-κB,p-NF-κB,IκB-α,and p-IκB-α protein expressions were significantly increased(P<0.05).Compared with LPS group,BV2 microglia intervened with different concentrations of DG had significantly higher viability(P<0.05),significantly lower NO secretion(P<0.05),significantly lower TNF-α,IL-6 and IL-1β inflammatory factors levels(P<0.05),significantly lower mRNA levels of TNF-α,IL-6,IL-1β,TLR4 and MyD88(P<0.05),and TLR4,MyD88,p-NF-κB and p-IκB-α protein expressions were significantly reduced(P<0.05).Conclusion:DG can inhibit LPS-induced neuroinflammatory responses in microglia,whose underlying mechanism is suppression of TLR4/NF-κB signaling pathway.

Objective To retrospectively analyze the diagnostic value of thrombin-antithrombin complex(TAT),thrombomodulin(TM),and tissue plasminogen activator-inhibitor complex(t-PAIC)in severe cases of corona virus disease 2019(COVID-19).Methods A cohort of 79 patients clinically diagnosed with COVID-19 was retrospectively assembled and categorized into two groups based on disease severity:non-severe(n=51)and severe(n=28).In this study the differences of coagulation function and inflamma-tory marker levels between the two groups were compared.The correlations of TAT,TM and t-PAIC with other biomarkers were investi-gated.The diagnostic values of all the markers for severe COVID-19 were assessed.Results The patients of severe COVID-19 exhibi-ted significantly higher levels of TAT,TM,and t-PAIC compared to those of non-severe group(P＜0.001).The levels of TAT,TM and t-PAIC showed notable positive correlation with other biomarkers.TAT demonstrated the strongest positive correlation with the level of D-dimer(r=0.786,P＜0.001).Binary logistic regression analysis identified TAT(OR=1.346,P＜0.05)and t-PAIC(OR=1.128,P＜0.05)were independent risk factors in term of severe COVID-19.The combined ROC curve for TAT,TM and t-PAIC revealed high diagnostic efficacy in severe cases with the area under the curve(AUCROC)were 0.918,and the sensitivity and specificity were of 75％and 94.1％,respectively.Conclusion The results of combined measurement of TAT,TM and t-PAIC effectively demonstrates its diagnostic value in identifying severity and stratification of COVID-19 cases and may have important clinical significance for assessment of the severity and prediction of the prognosis.

Objective To investigate the effect of different adhesive materials on all-ceramic restoration. Methods The all-ceramic restoration system model of mandibular first molar was established by spiral computed tomography (CT) scanning and computer-aided design (CAD) modeling. Four types of resin adhesive materials (Duo Cement,Lute-It,Rely-X ARC,Variolink II) used in clinics were selected, and the stress distribution was calculated using the ABAQUS software. Results The stress at the bonding interface was the highest when low filler adhesive Lute-It was used. Based on the viscoelasticity analysis, resin adhesives with a larger storage modulus and loss modulus could yield lower stress extremes. Conclusions The study suggests that high-filler type resin adhesives with a large energy storage modulus and loss modulus should be used clinically.

Objective To study the effect of Salidroside(Sal)on platelet activation and aggregation and the interaction between human platelets and MDA-MB-468 cells of breast cancer.Methods Human platelets were collected,platelet sus-pension was prepared,and platelets were treated with different concentrations of Sal.The effects of Sal on platelet activation and aggregation were detected by thromboelastogram(TEG)and flow cytometry.Breast cancer MDA-MB-468 cells were cul-tured in vitro,and human platelets were treated with different concentrations of Sal,and then activated by thrombin.The effects of Sal on the interaction between platelets and MDA-MB-468 cells were analyzed by adhesion test and scratch test.Re-sults TEG detection:The ADP inhibition rate in the blank control group was(10.97±12.69)%,and the ADP inhibition rate in all Sal intervention groups was higher than that in the blank control group(P＜0.05).The AA inhibition rate was(8.11±7.84)%in the control group and(25.96±15.18)%in the 5 μmol/L Sal intervention group,and the difference was statistically significant(P＜0.05).Flow cytometry:The expression of CD62P on platelet surface in 40 and 60 μmol/L Sal groups was(56.5±0.17)%and(65.50±0.36)%,respectively,and the difference was statistically significant compared with the positive control group(76.53±0.49)%(P＜0.05).The percentages of PAC-1 expression on platelet surface in 40 and 60 μmol/L Sal groups were(62.20±0.10)%and(58.47±0.15)%,and the difference was statistically significant com-pared with the positive control group(72.10±0.20)%(P＜0.05).Adhesion experiment:Platelets can have adhesion with MDA-MB-468 cells,and activated platelets have stronger adhesion ability.The adhesion rate in the Sal group was signifi-cantly lower than that in the positive control group,and was negatively correlated with the concentration of Sal.Scratch test:The cell mobility at 24 h in the positive control group was(12.71±0.70)%,and the cell mobility in each Sal treatment group was(4.51±0.44)%,(3.85±0.11)%,(5.37±0.36)%,(4.15±0.13)%and(3.55±0.38)%,respectively,showing significant decrease compared with the positive control group(P＜0.05).After 48 h of Sal treatment,the cell mobility of 10,20,40 and 60μmol/L Sal groups decreased,and there was a statistical difference compared with the positive control group(P＜0.05).Conclusion Sal can inhibit the adhesion between activated platelets mediated by thrombin and MDA-MB-468 cells and the migration of MDA-MB-468 cells.

Isopeptide bond-mediated molecular superglue is the irreversible covalent bond spontaneously formed by the side chains of lysine (Lys) and asparagine/aspartic acid (Asn/Asp) residues. The peptide-peptide interaction is specific, stable, and can be achieved quickly without any particular physicochemical factor. In the light of recent progress by domestic and foreign researchers, here we summarize the origin, assembly system and mechanism of isopeptide bond reaction, as well as the molecular cyclization and protein topological structure mediated by it. The prospect for its application in synthetic vaccine, hydrogel and bacterial nanobiological reactor is further discussed.
Cyclization ; Lysine ; Peptides ; chemistry ; Proteins

【Objective】 To investigate the feasibility of allogeneic platelet-rich plasma (PRP) for the treatment of herpes zoster wounds secondary to systemic lupus erythematosus (SLE), especially large ulcer wounds. 【Methods】 The treatment process of a patient with massive herpes zoster wounds in perineum and hip accompanied by extensive soft tissue necrosis secondary to SLE was retrospectively analyzed. The clinical efficacy of allogeneic PRP was explored combined with treatment key points and literature review. 【Results】 The patient′s wound bed was prepared until the wound was fresh, then treated externally with allogeneic PRP 3 times a week. The wound was healed completely after 42 days. 【Conclusion】 In the case of autologous PRP unavailable or unsuitable, allogeneic PRP is a safe alternative, which can effectively promote tissue regeneration, and this patient achieved curative effect in a short period of time.