OBJECTIVETo isolate and culture bovine cementum-derived cells and investigate their biological properties.

METHODSCulture of primary bovine cementoblast (CB) were established from new-born bovine teeth. Cementum was manually dissected, fragmented, and digested twice with collagenase. Following a thorough wash to remove liberated cells, the remaining cementum fragments were plated and cultured in RPMI1640 containing 10% FCS and 1.5% Hepes. The cells in culture were identified using immunocytochemistry by expression of cementum attachment protein (CAP), osteocalcin (OCN) and alkaline phosphatase (ALP). ALP activity was measured by modified Gomori staining.

RESULTS(1) The cells in culture possessed the typical morphology of CB and there were no obvious change of the cell morphology up to 5th passage; (2) Cells in culture exhibited alkaline phosphatase activity and were also positive to CAP, OPN and ALP immunohistochemistry staining.

CONCLUSIONThe cells isolated and cultured in this experiment possess the properties of CB. This enables us to further observe its biological characteristics during cementogenesis and periodontal regeneration.

Alkaline Phosphatase ; metabolism ; Animals ; Cattle ; Cell Separation ; methods ; Cells, Cultured ; Dental Cementum ; cytology ; enzymology ; Immunohistochemistry

OBJECTIVESTo study the essence of N-terminal amino acid sequence of 12 000-protein in gingival crevicular fluid (GCF).

METHODSGCF samples from patients with RPP and AP were collected. 12 000-protein was separated by SDS-PAGE and transformed to PVDF by electronic transformation. The aim band was cut to be analyzed in 491 Protein Sequencer.

RESULTSThe first ten of N-terminal amino acid sequence of 12 000-protein in GCF was Met, Leu, Thr, Glu, Leu, Glu, Lys, Ala, Leu, Asn. Through checking up in MS-Edman, the sequence was similar to "Ca binding protein, MRP8" which is the light subunit of Calprotectin.

CONCLUSIONSCalprotectin is a major protein in granulocytes and monocytes, and is related to many inflammatory diseases, maybe served as a effective marker for evaluating the inflammation of periodontium.

Amino Acid Sequence ; Electrophoresis, Polyacrylamide Gel ; Gingival Crevicular Fluid ; chemistry ; Humans ; Leukocyte L1 Antigen Complex ; Periodontium

OBJECTIVETo investigate the timing and location of the expression of bone sialoprotein (BSP) and osteopontin (OPN) in developing dental tissues of rats.

METHODSEvery three neonatal rats were sacrificed at day 1, weeks 1, 2, 3, 5 and 8. The mandibles were dissected and the first molar and the surrounding tissue were fixed, then demineralized with 15% EDTA. Immunohistochemical technique was used to determine the expression of BSP and OPN in dental tissues and the surrounding bone at different time points.

RESULTSImmunoreactivitis of BSP and OPN were present in matured ameloblasts, dentinoblasts, cementoblasts, osteoblasts, and the matrix. The expression of BSP and OPN in cementum and alveolar bone was stronger than that in enamel and dentine. In cementum and alveolar bone, BSP appeared to be concentrated in unmineralized and mineralized tissues, but OPN was concentrated in the mineralizing frontier and reversal line.

CONCLUSIONSBSP and OPN play an important role in the development and mineralization of rat mineralized tissues. The expression of BSP was different from OPN, indicating their different functions.

Animals ; Immunohistochemistry ; Integrin-Binding Sialoprotein ; Male ; Osteopontin ; Rats ; Rats, Sprague-Dawley ; Sialoglycoproteins ; analysis ; Tooth ; chemistry

Objective:To design and evaluate the application value of intracavitary-interstitial brachytherapy (IC-ISBT) applicator template for locally advanced cervical cancer.Methods:MRI data of 100 patients with ⅡB-ⅣA stage cervical cancer (International Federation of Gynecology and Obstetrics 2018 staging system) before and after external beam radiation therapy (EBRT) admitted to Sun Yat-sen University Cancer Center from March 2019 to September 2020 were collected. The range of primary cervical lesions was retrospectively analyzed and compared. Based on the residual mass of patients, the corresponding high-risk clinical target volume (HR-CTV) was delineated, and the IC-ISBT applicator template was designed and initially applied to cervical cancer patients. Dosimetry analysis and efficacy evaluation were compared between the applicator template-guided ( n=37) and free-hand implantation groups ( n=63). Chi-square test or Fisher exact test was performed for categorical variables, and t-test or U-test for continuous variables. Results:The median distance between the residual tumor margin (clockwise 3, 6, 9, 12 o'clock) and the center of 100 patients with ⅡB-ⅣA stage cervical cancer after EBRT was 16.5, 14.0, 17.0 and 13.0 mm, respectively. The corresponding HR-CTV was superimposed to reconstruct the three-dimensional diagram, and the cylindrical IC-ISBT applicator template with mushroom-like head was designed and manufactured: the longest and shortest diameter of the head was 35 and 20 mm, respectively; the central channel was adapted to the uterine tube, the C1-C12 channels was arranged in inner circle, and the peripheral B1-B5 and A1-A4 pin channels were expanded bilaterally. In terms of dose coverage, there was no significant difference between the HR-CTV D 90% [(635.12±22.65) vs. (635.80±25.84) cGy], bladder D 2 cm3 [(473.79±44.78) vs. (463.55±66.43) cGy)], rectum D 2 cm3 [(396.99±73.54) vs. (408.00±73.94) cGy] and sigmoid colon D 2 cm3 [(293.07±152.72) vs. (311.31±135.77) cGy] between the template-guided and free-hand implantation groups (all P>0.05), but the HR-CTV D 98% was significantly higher [(544.78±32.07) vs. (536.78±32.04) cGy, P=0.007] and the rectum D 1 cm3 and D 0.1 cm3 were significantly lower [(438.62±69.65) vs. (453.97±67.89) cGy, P=0.016; (519.46±70.67) vs. (543.82±81.24) cGy, P=0.001] in the template-guided implantation group. In addition, there was no significant difference in the complete response rate between two groups (86% vs. 83%, P>0.05). Conclusions:This IC-ISBT applicator template is reasonably designed, and the therapeutic efficacy of the template-guided implantation is equivalent to that of free-hand implantation. The dose coverage of the target area meets the clinical demand with a better protection of the organs at risk. The applicator template has the potential to be widely used as a conventional template in clinical practice as the applicator-guided implantation is convenient to operate and repeat.

OBJECTIVETo observe the efficacy of high-dose dexamethasone in combination with low-dose rituximab as a second-line treatment for patients with immune thrombocytopenia (ITP).

METHODS65 patients with ITP, previously by conventional dose of glucocorticoids, received high-dose dexamethasone in combination with low-dose rituximab (dexamethasone 40 mg/d for 4 days, rituximab 100 mg, d 7, 14, 21, 28 intravenous infusion). Treatment response, regulatory T cells (Treg), cytokines levels and treatment-related adverse effects were observed.

RESULTSTotal response rate 1 month after treatment was achieved in 81.5% (53/65) of patients, and complete response at 3,6 and 12 months was 72.3% (47/65), 66.2%(43/65), 63.1%(41/65). The higher efficiency and complete response rate was achieved in preexisting glucocorticoid-dependent patients. For patients with complete response, Treg cells continued to show a high level state [(3.01 ± 0.95)% vs (1.69 ± 0.35)%, P=0.032], cytokines of BAFF [(648.03 ± 79.63) ng/L vs (972.35 ± 93.64) ng/L, P=0.001], IL-2 [(2.84 ± 0.32) ng/L vs (4.18 ± 0.46) ng/L, P=0.012], sCD40L[(4.55 ± 0.66) ng/L vs (7.73 ± 1.04) ng/L, P=0.006] significantly lower than that before treatment. The level of IL-10 was increased, but without significance compared with that before treatment (P=0.136). All patients completed the protocol with no serious adverse reactions.

CONCLUSIONThe data show high-dose dexamethasone in combination with low-dose rituximab still has a satisfactory outcomes for patients previously with conventional dose of glucocorticoid.

Antibodies, Monoclonal, Murine-Derived ; Cytokines ; Dexamethasone ; Drug Combinations ; Glucocorticoids ; Humans ; Purpura, Thrombocytopenic, Idiopathic ; Rituximab ; T-Lymphocytes, Regulatory

OBJECTIVE: To analyze the predictors of successful weaning off extracorporeal membrane oxygenation (ECMO) after extracorporeal cardiopulmonary resuscitation (ECPR). METHODS: The clinical data of 56 patients with cardiac arrest who underwent ECPR in Hunan Provincial People's Hospital (the First Affiliated Hospital of Hunan Normal University) from July 2018 to September 2022 were retrospectively analyzed. According to whether ECMO was successfully weaning off, patients were divided into the successful weaning off group and the failed weaning off group. The basic data, duration of conventional cardiopulmonary resuscitation (CCPR, the time from cardiopulmonary resuscitation to ECMO), duration of ECMO, pulse pressure loss, complications, and the use of distal perfusion tube and intra-aortic balloon pump (IABP) were compared between the two groups. Univariate and multivariate Logistic regression analyses were performed to identify the risk factors for weaning failure of ECMO. RESULTS: Twenty-three patients (41.07%) were successfully weaned from ECMO. Compared with the successful weaning off group, patients in the failed weaning off group were older (years old: 46.7±15.6 vs. 37.8±16.8, P < 0.05), higher incidence of pulse pressure loss and ECMO complications [81.8% (27/33) vs. 21.7% (5/23), 84.8% (28/33) vs. 39.1% (9/23), both P < 0.01], and longer CCPR time (minutes: 72.3±19.5 vs. 54.4±24.6, P < 0.01), shorter duration of ECMO support (hours: 87.3±81.1 vs. 147.7±50.8, P < 0.01), and worse improvement in arterial blood pH and lactic acid (Lac) levels after ECPR support [pH: 7.1±0.1 vs. 7.3±0.1, Lac (mmol/L): 12.6±2.4 vs. 8.9±2.1, both P < 0.01]. There were no significant differences in the utilization rate of distal perfusion tube and IABP between the two groups. Univariate Logistic regression analysis showed that the factors affecting the weaning off ECMO of ECPR patients were pulse pressure loss, ECMO complications, arterial blood pH and Lac after installation [pulse pressure loss: odds ratio (OR) = 3.37, 95% confidence interval (95%CI) was 1.39-8.17, P = 0.007; ECMO complications: OR = 2.88, 95%CI was 1.11-7.45, P = 0.030; pH after installation: OR = 0.01, 95%CI was 0.00-0.16, P = 0.002; Lac after installation: OR = 1.21, 95%CI was 1.06-1.37, P = 0.003]. After adjusting for the effects of age, gender, ECMO complications, arterial blood pH and Lac after installation, and CCPR time, showed that pulse pressure loss was an independent predictor of weaning failure in ECPR patients (OR = 1.27, 95%CI was 1.01-1.61, P = 0.049). CONCLUSIONS Early loss of pulse pressure after ECPR is an independent predictor of failed weaning off ECMO in ECPR patients. Strengthening hemodynamic monitoring and management after ECPR is very important for the successful weaning off ECMO in ECPR.
Humans ; Extracorporeal Membrane Oxygenation ; Blood Pressure ; Retrospective Studies ; Perfusion ; Cardiopulmonary Resuscitation