Objective To investigate the the multi-directional differentiation potential between pluripotent of human gingival fibroblasts (HGFs) and human periodontal ligament cells (HPDLCs). Methods HPDLCs and HGFs were obtained from the primary culture. HPDLCs and HGFs at 3rd-4th passage were cultured in osteogenic, adipogenic or chondrogenic me-dium. Cells without differentiation were taken as control. Alizarin red, Alcian blue and oil red O staining were performed to detect osteogenic differentiation, chondrogenic and adipogenic differentiation in vitro, respectively. Reverse transcription polymerase chain reaction (RT-PCR) was applied to examine the expression of osteocalcin (OCN), runt-related transcription factor 2 (RUNX2) and collagen 1 (Col 1), peroxisome proliferator-activated receptor gamma 2 (PPARγ2) and collagen 10 (Col 10). Results HPDLCs and HGFs cultured in osteogenic medium showed massive calicium nodulus at day 28, but HP-DLCs formed more calicium nodulus than those of HGFs. The expressions of OCN, RUNX2 and Col 1 were significantly high-er in HPDLCs than those in HGFs (P<0.05). In chondrogenic medium both cells were found blue deposit at day 14, and the expression of Col 10 was significantly higher in HGFs than that of HPDLCs (P<0.01). Furthermore, in adipogenic medium HGFs showed more lipid-filled droplets stained with oil red O than HPDLCs at day 21. The expression of PPARγ2 was sig-nificantly higher in HGFs than that of HPDLCs (P<0.01). Conclusion HPDLCs has the better potency of osteogenic differ-etiation than HGFs, however, HGFs has the better potency of adipogenic and chondrogenic differentiation.

Objective To investigate the pluripotency of human gingival fibroblasts (hGFs), and provide a novel cell source for tissue engineering. Methods With informed consent from volunteers, fresh and healthy gingiva were collected. The hGFs were obtained from the gingiva by tissue culture. The third passage of hGFs was cultured in osteogenic medium, chondrogenic medium and adipogenic medium. Cells without differentiation were taken as control. Cells were examined by al?kaline phosphatase (ALP) staining, Alizarin red staining, Alcian blue staining and oil red O staining for detecting of the abili?ty of differentiation pluripotency. Real-time polymerase chain reaction was applied to examine the expression of osteogenic marker genes ALP, runt-related transcript factor 2 (Runx2), chondrogenic marker aggrecan (AGR) and adipogenic marker peroxisome proliferator-activated receptor gamma 2 (PPARγ2). Results The hGFs cultured in osteogenic medium showed massive violet deposit at day 7 and calcium nodulus at day 28, meanwhile, the expressions of ALP and Runx2 were higher than those of control (P<0.01). In chondrogenic group cells were found blue deposit at day 14. In adipogenic group lipid-filled droplets stained with oil red O were found in cells at day 14. However, hGFs in control group had no any positive stain?ing. Furthermore, expressions of AGR and PPARγ2 were significantly higher than those of control (P<0.01). Conclusion Human gingival fibroblasts have the pluripotency of osteogenic, adipogenic and chondrogenic differentiation.

Objective:To compare the effectiveness between three methods for purifying the immunoglobulin of egg yolk(IgY) which are polyethylene glycol (PEG) method, chloroform extraction method and chloroform / PEG method, and to provide basis for obtaining the batch of IgY.Methods:The inactivated vaccine of Vibrio parahemolyticus (V. parahemolyticus) was prepared and the hens were immunized by multi-point intramuscular injection.The eggs were collected and the IgY was purified by PEG method, chloroform extraction method and chloroform/PEG method.The protein extraction rate, the IgY titer and the purity of the antibody which purified by different methods were detected.Furthermore, the operation process, cost and safety of the three methods were analyzed.Results:The protein contents of the extraction belonging three methods from high to low in turn were chloroform extraction method, chloroform/PEG method, and PEG method.There was no significant difference in the antibody titer between three methods, and the tiler of chloroform extraction method was slightly high.The purities of purified antibody from high to low in turn were PEG method, chloroform/PEG method and chloroform method.The PEG method had better security but relatively lower extraction efficiency and higher cost.The chloroform/PEG method had high extraction efficiency and good antibody purity.Conclusion:The PEG method is suitable for a small amount of extraction in the laboratory.The chloroform/PEG method is appropriate for extracting the high quality IgY in a batch as it has high extraction efficiency and good antibody purity.

The lipase labeled with the fluorescein isothiocyanat (FITC) was immobilized on the derivatives of the polyethylene glycol. The article discussed the effect of factors on the characters of lipase and analyzed the relationships among the activity of lipase, conformation, and fluorescence spectrum while the activity and the fluorescence spectrum of immobilized lipase were determined. The results demonstrated that polyethylene glycol 400-diacrylate could form appropriate network to improve the activity of enzyme. Adding ligand induced the lipase's catalytic conformation to increase the activity twice more than before. The active centre of lipase could be released by the extraction of ligand thus increasing the activity. After immobilization, the stability of labeled lipase improved greatly: immobilized lipases retained more than 70% and 60% of initial activity under conditions of 90 degrees C and strong acid or alkali, respectively. After immersing immobilized lipases into guanidine hydrochloride or urea for 15 days, the lipases retained upwards of 70% activity. The fluorescence spectrum could obviously reflect the changes of the activity and conformation of lipase. The fluorescence intensity was the minimum in the optimal pH and temperature. In the denaturing agent it declined as time passed. These results indicated that the unfolded processes of immobilized lipases are different under different conditions.
Dextrans ; chemistry ; Enzyme Stability ; Enzymes, Immobilized ; chemistry ; metabolism ; Fluorescein-5-isothiocyanate ; analogs & derivatives ; chemistry ; Fluorescent Dyes ; chemistry ; Lipase ; chemistry ; metabolism ; Polyethylene Glycols ; chemistry ; Protein Unfolding ; drug effects

OBJECTIVE: To investigate the diagnostic value of transcranial Doppler ultrasound in early stage cerebral arteriosclerosis. METHODS: We selected 50 patients with early cerebral arteriosclerosis as the disease group. At the same time we selected another 50 patients as a control group with no significant symptoms in the nervous system. By 2 MHz pulse Doppler probe through double-temporal windows and pillow windows Basilar artery (BA), the bilateral middle cerebral artery (MCA) was detected. In the TCD spectrum, we selected the spectrum of a single-family cardiac cycle, identified the starting point (ts), pulse-incisure point (ti), end point (te), and the peak velocity (tp), measured the time of the spectrum starting point to the peak velocity (Tp) and calculated the time required for the peak velocity in the share of ventricular systolic (Tp/Ti), the time required for the peak velocity in the share of the whole cardiac cycle (Tp/T). Tp, Tp/Ti and Tp/T were respectively named as time to peak velocity (TPV), peak-time index-1 (PTI-1) and peak-time index-2 (PTI-2). All data were analyzed by SPSS13.0. RESULTS: There was no significant difference in blood vessel velocity, PI, RI and S/D of BA and RMCA (P>0.05) between the control group and the disease group. Compared with the control group, TPV of the BA, LMCA and RMCA significantly extended, PTI-1 and PTI-2 of BA, LMCA and RMCA increased significantly in the disease group (P<0.01). In the disease group, there was no significant correlation between peak time index and PI, S/D (P>0.05). CONCLUSION TPV, PTI-1 and PTI-2 are sensitive indicators of early stage cerebral arteriosclerosis.
Adult ; Basilar Artery ; diagnostic imaging ; Blood Flow Velocity ; physiology ; Case-Control Studies ; Female ; Humans ; Intracranial Arteriosclerosis ; diagnostic imaging ; Male ; Middle Aged ; Middle Cerebral Artery ; diagnostic imaging ; Ultrasonography, Doppler, Transcranial

Objective:To investigate the perception status of inpatients to nurses' caring behavior, and analyze the correlation between them and nurse-patient relationship trust degree. Methods:Adopting general information questionnaire, Caring Behaviors Inventory ( CBI) , nurse-patient relationship trust scale, a questionnaire survey was conducted among 226 inpatients in a third class A tertiary hospital in Tianjin. Results:The total score of ca-ring behavior perception was (96. 92 ± 15. 68), and among the four dimensions, the perception of patients to nur-ses' knowledge and skills was deepest, the perception to nurse' s contact with patients was worst; total score of nurse-patient trust scale was (124. 75 ± 19. 13). There was a positive correlation between the total score of pa-tients' caring perception and nurse-patient relationship trust degree (r=0. 554);multiple linear stepwise regres-sion analysis showed respect for patients, support and assurance were the main factors influencing the nurse-pa-tient relationship trust degree. Conclusion:The patients' perception on nurses' caring behavior and nurse-pa-tient relationship trust degree is closely related. Nurses should carry out targeted care to patients with different char-acteristics to improve patients' trust to nurses and construct a harmonious nurse-patient relationship.

Objective To investigate the effects of risk factors on non thyroidal Illness syndrome (NTIS) in elderly inpatients.Methods A total of 819 elderly inpatients who met inclusion criteria were consecutively recruited in thiscross-sectional study.Physicalmeasurements and mini nutritional assessment using the mini nutritional assessment-short form (MNA-SF) score were conducted.A serum levels of thyroid stimulating hormone (TSH),free triiodothyronine (FT3),free thyroxine (FT4),brain natriuretic peptide (BNP) and C-reactive protein (CRP) were examined.Data were analyzed with multivariatelogistic regression.Results The significant differences were found between NTIS group (n=145) versus control group (n=674)inage (78.5±8.1) years vs.(75.1±8.6) years(t=5.422,P＜0.01),in body mass index (23.0 ±3.8) kg/m2 vs.(24.1±3.6) kg/m2,in MNA-SF score 11.2±2.3 vs.12.3± 1.8(t=-3.315,6.754,P＜0.01),in level of serum albumin (36.0±4.5) g/L vs.(38.4±3.6) g/L (t=-6.977,P＜0.01),in triglyceride level (1.3± 0.9) mmol/L vs.(1.5±1.0) mmol/L(t=-3.039,P＜0.01),inCRP (Z=-8.857,P＜0.01)),and in BNP (t=6.331,P＜0.01).Logistic regression analysis revealed that age＞ =80 years (OR=2.433,95％CI:1.357 4.361),malnutrition (OR=1.946,95％CI:1.261-3.001),renal insufficiency (eG FR＜60 ml/min,OR =2.131,95％ CI:1.367-3.322),and high level of CRP (10 mg/L and 50 mg/L:OR=3.446,95％CI:2.117-5.611;over 50 mg/L:OR =10.029,95％CI:4.693-21.432,all P＜0.01)) were risk factors for NTIS.Conclusions Non-thyroidal illness syndrome in elderly inpatients is correlated with advanced age,renal insufficiency,malnutrition and stress,which are the independent risk factors.

Since the end of 2019, the COVID-19 caused by 2019-nCoV has emerged and the pandemic ravaged the world, which seriously threatens global public health security and economic development. 2019-nCoV vaccine is an effective weapon to combat the viral infection, however, studies have shown that vaccine-induced immune protection decreases over time, coupled with some novel and immune escape variants continual emerging. Therefore, it is urgent to complete booster immunization to improve protection. At present, 2019-nCoV vaccines based on a variety of technical platforms have been approved and available in China. Therefore, we developed this sequential vaccination strategy guide to provide documentation guidance for the prevention and control of the epidemic caused by 2019-nCoV and its variant strains.

Objective:To identify the risk factors for massive blood transfusion in pediatric living donor liver transplantation.Methods:The medical data of children underwent living donor liver transplantation in our hospital from April 2006 to April 2019 were retrospectively collected.Massive transfusion was defined as the administration of red blood cells > 1 fold of the total blood volume (70 ml/kg) during operation.Patients were assigned to massive transfusion group and non-massive transfusion group according to the volume of blood transfused during operation.Binary logistic regression analysis was used to identify the risk factors for massive blood transfusion during living liver transplantation.Results:A total of 95 pediatric patients were enrolled in this study, with 18 cases in massive transfusion group and 77 cases in non-massive transfusion group.The incidence of massive blood transfusion was 19% during operation.The results of logistic regression analysis showed that preoperative survival status of " hospitalization" ( OR=49.816, 95% CI 2.945-842.59, P=0.007), increased serum Cr concentrations ( OR=1.046, 95% CI 1.007-1.086, P=0.021), increased Pediatric End-Stage Liver Disease (PELD) or Model for End-Stage Liver Disease (MELD) score ( OR=1.215, 95% CI 1.046-1.411, P=0.011) and prolonged operation time( OR=1.623, 95% CI 1.133-2.327, P=0.008) were the independent risk factors for intraoperative massive blood transfusion in living donor liver transplantation, while increased recipient weight ( OR=0.856, 95% CI 0.761-0.962, P=0.009) was a protective factor for intraoperative massive blood transfusion. Conclusions:Preoperative survival status of " hospitalization", increased PELD or MELD score and prolonged operation time are independent risk factors, while increased pediatric weight is a protective factor for massive blood transfusion in pediatric living donor liver transplantation.

Objective:To explore the effects of microRNA-126 (miR-126) on the proliferation of human myeloid leukemia mononuclear cells (THP-1)-derived macrophages in high glucose environment and the regulatory role of miR-126 in periodontitis with diabetes.Methods:THP-1 cells were cultured in vitro and 5 μg/L phorbol-12-myristate-13-acetate was applied to induce THP-1 cells differentiating into macrophages for 48 h in low glucose culture medium (5.5 mmol/L). THP-1-derived macrophages were then cultured with low glucose, medium glucose (15 mmol/L) or high glucose (25 mmol/L) media respectively. The proliferation of THP-1-derived macrophages was detected by cell counting kit-8 (CCK-8) method and the expressions of miR-126 and proliferation-associated factors were detected by quantitative real time PCR (qRT-PCR). The miR-126 mimic or inhibitor was transfected into THP-1-derived macrophages for 72 h. The proliferation of cells was detected by CCK-8 method and the expressions of miR-126 or proliferation-associated factors were detected by qRT-PCR. Results:Increasing glucose concentration decreased the proliferation of THP-1-derived macrophages (day 7, A values in low, medium and high glucose groups were 0.369±0.014, 0.214±0.009 and 0.200±0.010, respectively, P<0.01) as well as the survival rate ( P<0.05), promoted the expression of miR-126, B-cell lymphoma-2 (Bcl-2)-associated X protein (BAX) and caspase-3 ( P<0.05), and suppressed Bcl-2, phosphoinositol-3 kinase regulatory subunit 2 (PIK3R2) expression ( P<0.05). After the miR-126 mimic was transfected in cells in low glucose medium for 72 h, compared with negative control (1.005±0.118), the expression of miR-126 significantly increased (2 980.227±170.431, P<0.05), and the proliferation of THP-1 derived macrophages decreased (negative control: 1.816±0.013, mimic group: 1.310±0.048, P<0.01), the level of BAX and caspase-3 significantly increased ( P<0.01, P<0.05), PIK3R2 and Bcl-2 significantly decreased ( P<0.05, P<0.01). After the miR-126 inhibitor was transfected in cells cultured in high glucose medium for 72 h, compared with negative control (0.723±0.133), the proliferation of inhibitor group increased (0.984±0.049, P<0.05), the level of BAX and caspase-3 significantly decreased ( P<0.01, P<0.05), PIK3R2 and Bcl-2 significantly increased ( P<0.01, P<0.05). Conclusions:High glucose condition can inhibit the proliferation of THP-1-derived macrophages and increase the expression of miR-126. MiR-126 can inhibit the proliferation of THP-1-derived macrophages in high glucose environment through up-regulating the expression of BAX and caspase-3 and down-regulating the expression of PIK3R2 and Bcl-2.