OBJECTIVEThis study explores the effects of different fibrin glue combination modes on the survival, proliferation, and apoptosis of dental follicle cells (DFCs), as well as to evaluate the feasibility and effectiveness of fibrin glue as transplantation material.

METHODSThe membranes of surviving DFCs were marked using 3,3'-dioctadecyloxa carbocyanine perchlorate (DIO), and the cell number was counted by using ImageJ2x software. The apoptotic cells were marked with prodium iodide (PI).

RESULTSCompared with that of the 3D-2 and 2D-1 groups, the degradation speed of the 3D-1 group was the slowest. DFCs could survive and grow well in fibrinogen with a concentration of 15 mg · mL⁻¹ supplemented with thrombin with a concentration of 2 U · mL⁻¹. In particular, the 3D-1 combination mode was significantly conducive to cell proliferation and stretching.

CONCLUSIONFibrin glue can be used as an effective cell transplantation material. The different combination modes have certain effects on cell proliferation. The 3D-1 combination mode is more conducive to the survival and proliferation of DFCs than other modes.

Apoptosis ; Cell Proliferation ; Cell Survival ; Dental Sac ; cytology ; Fibrin Tissue Adhesive ; pharmacology ; Fibrinogen ; Humans ; Thrombin

Objective To evaluate the effect of ecoimmunonutrition on systemic inflammatory response in swine with acute necrotizing pancreatitis (ANP). Methods Swine model of ANP was induced by retrograde injection of 5% sodium taurocholate and trypsin into the pancreatic duct. 24h after the ANP models were established, eighteen swine were randomly divided into total parenteral nutrition (PN) group (n = 6), enteral nutrition (EN) group (n = 6), and ecoimmunonutrition (E1N) group (n = 6). Each of the three nutrition formulas was given to each group respectively for eight days. The level of plasma endotoxin and serum concentrations of TNF-α, IL-1β, IL-6 and IL-10 were measured before, 24 h after the induction of ANP and 1,2, 4, 8 d after nutrition support. Results 24 hours after the ANP models were established, the serum level of plasma endotoxin, TNF-α, IL-1β, IL-6 and IL-10, NF-κB activities in EIN group were ( 1.85 ± 0.18) EU/L,(461.59 ± 128.25 ) pg/ml, ( 185.49 ± 58.81 ) pg/ml, ( 354.26 ± 34.63 ) pg/ml, ( 110.32 s 25. 18 ) pg/ml,(51.06 ± 2.27 )%, respectively, which all were significantly higher than those before the ANP models were established, but there were no difference among three groups. Eight days after nutrition support, those levels in EIN group were (1.48 ±0.16 )EU/L, (30.11 ±9.12)pg/ml, (20.17 ±7.04)pg/ml, (36.42 ±7.24) pg/ml and ( 89.46 ± 13.54 ) pg/ml, (9.06 ± 0. 17 ) %, respectively, which all were significantly lower than those of TPN and EEN group ( P ＜ 0. 05 ). The ration of IL-10/IL-6 was 2.51 ± 0.42 in EIN group, and 2.28 ± 0. 19 before the ANP models were established. Conclusions EIN could attenuate endotoxemia, decrease NF-κB activities and serum concentrations of cytokines and maintained the balance of pro- and anti-inflammation.

Objective To evaluate the effect of ecoimmunonutrition on maintenance of the mucosal barrier function in pigs with severe acute pancreatitis(SAP).Methods Twenty pigs were injected with5% sodium taurocholate and trypsin into the pancreatic duct to establish SAP models.Twenty four hours later,18 suvival pigs were randomly given parenteral nutrition(PN group,n=6),elemental enteral nutrition(EEN group,n=6),or ecoimmunonutrition(EIN group,n=6)for 8 days.The serum amylase,endotoxin and intestinal permeability were determined on different times.All pigs were sacrificed 8days later,and their venous blood,mesenteric lymph nodes(MLN),liver,lung,spleen and pancreas specimens were cultured quantitatively and qualitatively with standard techniques.The morphology of small bowel was observed and pathological changes of the pancreas was analyzed.Results Serum amylase of all groups were obviously elevated,but there was no significant difference among them(P＞0.05).The plasma endotoxin and intestinal permeability were lower in EIN group[(1.50±0.18)EU/L and(42.8±20.0)×10-3 respeetively]compared to EEN group[(1.98 4±0.20)EU/L and(67.4±23.0)×10-3 respectively]and PN group[(3.96±0.40)EU/L and(197.2±47.4)×10-3.respectively](all P value＜0.05).The incidence and the magnitude of bacterial translocation in EIN group were lower than those in PN and EEN groups.However,the ileum mucosa thickness,villus height,crypt depth and the rate of the normal intestinal villi in EIN group were(398.27±52.93)μm,(269.72±41.66)μm,(681.98±58.33)μm and 79%,respectively,but in PN group were(218.32±35.81)μm,(145.76±23.34)μm,(376.20±48.23)μm and13%,respectively and in EEN group were(305.70±42.72)μm,(192.52±38.17)μm,(507.31±68.23)μm and 47%,respectively.Conclusions EIN can maintain the integrity of intestinal mucosal barrier and decreasing intestinal bacterial and endotoxin translocation in SAP pigs.

Objective To explore the clinical value of high‐sensitive troponin I detection in the diagnosis of early myocardial inju‐ry .Methods Totally 240 type 2 diabetes mellitus (T2DM ) patients (study group) and 40 healthy people(healthy control group) were chosen as subjects .The serum level of HbA1c ,AST ,CK ,CK‐MB and hs‐cTnI were measured among the two groups ,respec‐tively .Results Hs‐cTnI level in T2DM group was significantly higher than in healthy control group(P< 0 .05) ,and AST ,CK and CK‐MB levels in two groups had no significant difference(P> 0 .05) .Hs‐cTnI level and positive rate in HbA1c ≥ 6 .5% group was significantly higher than in HbA1c ≤ 6 .4% group(P< 0 .05) ,and no significant difference of AST ,CK and CK‐MB levels and posi‐tive rate were observed in two groups(P> 0 .05) .There was a positive correlation between the level of HbA1c and that of serum hs‐cTnI in T2DM group(r= 0 .471 ,P< 0 .05) .Conclusion The hs‐cTnI level detection has important clinical value in the diagnosis of early myocardial injury in T2DM patients ,and HbA1c is positively correlated with hs‐cTnI .

Objective To explore the diagnostic value of five infection markers in bloodstream infection.Methods Randomly selected 110 bloodstream infection patients with positive blood cultures and 30 bacterial infection patients with negative blood cultures.Blood was simultaneously drawn with blood cultures;the complete blood count and C-reactive protein (CRP) levels were measured.The white blood cell count (WBC),neutrophil count (NEU),lymphocyte count (LMY),CRP level and neutrophil-lymphocyte count ratio (NLCR) were compared between the two groups.Results The levels of WBC,NEU,NLCR and CRP in bloodstream infection group were significantly higher than those in control group (P＜0.05),while LYM was significantly lower than that in control group (P＜ 0.05).Among these five infection markers,the area under the receiver operating characteristic curve (ROC-AUC) was the highest for NLCR (0.808) and LMY (0.756);when the cutoff value for NLCR was ＞9.33,sensitivity was 63.6％,specificity was 93.3％;and the cutoff value for LYM was ≤0.97,sensitivity was 58.2％,specificity was 86.7％.Furthermore,the NLCR of patients with gram-negative bloodstream infection was higher than those in patients with gram-positive bloodstream infection.NLCR showed important clinical significance in distinguishing strains of different bloodstream infections.Conclusions NLCR is the better predictors than routine parameters in diagnosing bloodstream infection.

BACKGROUND:Titanium aloy and stainless steel are two common internal fixation materials, but there are some difference in their therapeutic effects and biocompatibility. OBJECTIVE:To explore the therapeutic effects and biocompatibility of titanium aloy and stainless steel internal fixation materials for the treatment of spinal tuberculosis. METHODS:Seventy-one spinal tuberculosis patients, 35 males and 36 females, aged 17-81 were enroled. Among them, 35 patients received titanium aloy internal fixation, and the 36 patients underwent stainless steel internal fixation. At the end of 12-month folow-up, Cobb angle changes, therapeutic effect and Frankel grade were analyzed in the two groups. RESULTS AND CONCLUSION:Before treatment, there was no difference in the spinal kyphosis angle and Frankel grade between the two groups. At the last folow-up, the Frankel grade and Cobb angle were both improved in the two groups (P < 0.05), but there was stil no difference between the two groups. The cure rate was 97% (n=34) in the titanium aloy group and 92% in the stainless steel group (n=33), and no significant difference was found between the two groups. These two kinds of internal fixation materials exhibited good biocompatibility, and no infection and other adverse reactions occurred. These findings indicate that both titanium aloy and stainless steel as internal fixation materials have good biocompatibility and therapeutic effects.

Objective To investigate the expression of receptor tyrosine kinase EphB2 and its relationship with peritumoral brain edema (PTBE) in meningiomas and to explore the possible mechanism of PTBE in meningiomas. Methods A total of 44 meningioma tissue samples from patients undergoing surgery and 14 normal meninges samples as the control were collected for the examination of the immunoreactive EphB2 expression. The expressions of EphB2 in different types of meningiomas were determined by immunohistochemical method. Peritumoral brain edema was assessed on a subjective 5 point(0～4) scale based on the data from preoperative magnetic resonance imaging scan. Results ① The expressions of EphB2 in 44 cases of meningiomas were: scale 4 in 7 cases, scale 3.5 in 5 cases, scale 3 in 6 cases, scale 2.5 in 2 cases, scale 2 in 8 cases, scale 1 in 13 cases, scale 0 in 3 cases, mean scale 2.19 . ② No expression of EphB2 was found in 10 out of the 14 normal meninges samples, but minimal (scale 1) expression of EphB2 was found in only 4 cases. ③ Preoperative MRI results showed different levels of PTBE in 33 out of 44 cases. Conclusion There is expression of EphB2 in the cytoplasm in most meningiomas. The expression of EphB2 in meningnioma is positively correlated with peritumoral brain edema, suggesting that EphB2 participates in PTBE of meningiomas.

Objective To evaluate the diagnostic value of bedside video electroencephalogram (VEEG) in neonatal within 6 h after birth in diagnosing hypoxic-ischemic encephalopathy ( HIE) and the correlation of bedside VEEG results and early neural and behavioral development. Methods Neonatal severe asphyxia cases were collected and bedside VEEG and HIE were recorded and graduated. The sensitivity and specificity of different VEEG degrees within 6 h after birth were calculated in prognosing HIE degree. The sensitivity and specificity of abnormal VEEG performed within 6 h, the third day and the seventh day after birth, respectively, were compared in prognosing HIE. Neonatal behavior neurological assessment (NBNA) was performed at 7 to 14 days of age, EEG and general movements assessment (GMs), developmental screening test for child under six(DST) when patients were 3 months old, and EEG, Bayley scales of infant development(BSID) at 6 months old to analyze the correlation of bedside VEEG results and early neural and behavioral development. Results Forty-eight severe asphyxia neonatal were included, among which 12 severe asphyxia and 36 HIE, including 14 mild, 12 moderate and 10 severe HIE. There were nine normal and 39(81. 3%) abnormal VEEG including 16 mild, 11 moderate, five severe abnormal and seven inactive VEEG within 6 h after birth. There were 32(88. 9%) abnormal VEEG within 6 h after birth in 36 HIE patients. Significant positive correlation was found between VEEG within 6 h after birth and HIE (r= 0.849, P<0. 01). Severe abnormal and inactive EEG within 6 h after birth showed sensitivity of 100%, specificity of 94. 7% in predicting severe HIE. The sensitivity of VEEG testing at 6 h,3 and 7 d in predicting HIE were 88. 9%, 83. 9% and 28. 6% , correspondingly the specificity were 41.7%, 91. 7% and 100%, respectively. Nine patients with continually abnormal VEEG died in hospital. The NBNA scores of patients with moderate and severe abnormal EEG were significantly lower than those with normal EEG (both P<0. 01), the NBNA scores of patients with severe abnormal EEG were significantly lower than those with moderate abnormal EEG (P<0. 05). Thirty-five patients were followed up in the hospital at 3 months old, and 32 patients had DST >85 and three had DST between 70 and 84 with abnormal EEG. GMs assessment of one of the three patients showed absence of fidget movements, cuing a chance of cerebral palsy. Fourteen patients were followed up in hospital at 6 months old, and seven of them had abnormal EEG, four had abnormal BSID with abnormal EEG. Conclusions VEEG within 6 h after birth shows high sensitivity and specificity in prognosing HIE, and much relates to short-term neural and behavior development.

Child ; Female ; Gastric Mucosa ; pathology ; Gastritis, Hypertrophic ; diagnosis ; Humans ; Hypertrophy ; diagnosis

Background Retinal pigment epithelium (RPE) cell transplantation is the primary means of human trial for the treatment of retinal degeneration.Induced pluripotent stem cells (iPSCs) will become an important source for cell transplantation.In addition,iPS-RPE cells may provide a personalized treatment platform for the patient's own cells treatment.Objective This study was to evaluate the feasibility of human fibroblasts differentiate toward iPSCs from retinitis pigmentosa (RP) patients and toward iPSC-RPE cells from non-RP individual by retroviral transfection of Oct4,Sox2,c-Myc and KLF4 genes.Methods Human thigh skin tissues were obtained from a RP patient with hotspot mutation of SNRNP200 p.S1087L and individual without SNRNP200 p.S1087L mutation,respectively,with the size 2 c m×3 cm.Human dermal fibroblasts were isolated and cultured by trypsin digestion and explant method.The fibroblasts were transfected by a series of retrovirus and cultured by human embyonic cellconditioned medium to generate and induce iPSCs,and then the iPSCs were identified by morphology,alkaline phosphatase (AP) staining and immunofluorescence assay of specific markers of pluripotent stem cells.iPSCs suspension were injected into SCID mouse to observe the tumorigenesis.The iPSCs from non-RP subject were induced to differentiate toward iPS-RPE cells by embryonic body (EB) inducing method,and iPS-RPE cells were identified by detecting the expression of RPE65,zonula occludens protein 1 (ZO-1) and lecithin retinol acyltransferase (LRAT).Results Cultured human dermal fibroblasts showed fusiform or polygon shape and intercellular close arrangement,and Vimentin was positively expressed in the cells.Small cell colonies were harvested 5-7 days after infected by retroviruses,and the morphology changed from spindle into round mass.The hESC-like iPSCs clonies appeared 20 days after cultivation,and the positive expressions of hESC-specific surface antigens including SSEA3,SSEA4,TRA-1-60,TRA-1-81 and Nanog were found in the cells 25-30 days after cultivation,and the positive staining of AP was obtained 12 weeks after cultivation.A teratoma was formed at the injection site of iPSCs suspension in SCID mouse.Immunofluorescence technique showed that RPE cell-specific proteins including RPE65,ZO-1 and LRAT proteins were positively expressed in iPS-RPE cells at 30 days after differentiation.Conclusions Mutation SNRNP200 p.S1087L of RP patient-specific iPSCs can be induced from human dermal fibroblast by retrovirus infection method.The function and morphology of the iPSCs were similar to hESCs.Human iPSCs cell line generated from the dermal fibroblasts of non-RP individuals can differentiate into iPS-RPE cells.