Bone Transplantation ; Dental Research ; Humans ; Tooth

Objective To explore the efficacy of multilevel anterior cervical discectomy and fusion for aged patients with cervical spondylotic myelopathy. Methods 25 aged patients with cervical spondylotic myelopathy were treated by multilevel anterior cervical discectomy and fusion. Japanese Orthopedic Association (JOA) scores were evaluated,clinical results were graded from excellent to poor using Odoms criteria,and the sagittal alignment of fusion segements were measured using Cobb method. Bone fusion and complications was evaluated. Results The final follow up score according to Odoms criteria was excellent in 8 patients ,good in 14 patients,and fair in 3 patients ,with excellent and good rate of 88%. The JOA score at final follow up(7.8) was significantly higher than preoperative(12. 8),with average improvement rate of 54%. At final follow up,bone fusion rate was 100%. Conclusion Multilevel anterior discectomy and fusion for aged patients with cervical spondylotic myelopathy was effective.

Objective To obtain alpaca single domain antibody targeting Her2.Methods An alpaca was immunized with human recombination Her2 protein mixed with Freund's adjuvant.Total RNA was extracted from the alpaca's blood and was used to synthesize first strand cDNA.Single domain antibody variable region (VHH) gene of the alpaca was amplified by PCR and cloned into pMES4 vector for library construction.After screening, E.coli BL21 (DE3) was transformed with selected clones and was induced with IPTG for the expression of recombinant proteins.The nanobody was purified by nickel ion affinity chromatography column.The affinity of the nanobodies to Her2 was tested.Results After the second round of screening, two antibody clones were selected, H3 and H5.The affinity of H5 was 8.106×10-10mol/L.Histochemistry results showed that H5 could recognize Her2 antigen in breast tumor tissue.Conclusion An Her2 specific nanobody derived from alpaca is obtained through phage display library screening, which can recognize human Her2 antibody in human breast tumor tissue.

BACKGROUND:Stable and efficient labeling of dental pulp stem cel s in vitro is most important in tracer technique, which is also the basis of tooth regeneration in vivo.
OBJECTIVE:To determine the optimal condition and method for transfection of stem cel s derived from rat dental pulp with green fluorescent protein infection by lentiviral vector and to determine whether green fluorescent protein-labeled dental pulp stem cel s maintain their stem cel properties.
METHODS:Rat dental pulp stem cel s were obtained by modified enzyme digestion method, to identify the immune phenotype and differentiation potential. Dental pulp stem cel s were infected with green fluorescent protein by lentiviral vector for 24 and 48 hours at different multiplicity of infection (MOI) (5, 10, 25, 50 and 100). The infection efficiency and fluorescence intensity were analyzed by inverted fluorescent microscopy. The clonal and proliferation ability, cel cycle and the mineralization potential were compared before and after transfection. Based on those mentioned above, we could evaluate the influence of infection on their biological characteristics.
RESULTS AND CONCLUSION:Flow cytometry results showed that rat dental pulp stem cel s expressed STRO-1 and CD146 but not CD34 or CD45. The dental pulp stem cel s could differentiate into osteoblasts and
adipocytes when cultured in specific medium for each lineage differentiation. The highest efficiency of infection and strongest fluorescence expression appeared at 48 hours of infection and MOI 50. There were no significant differences in growth ability, cel colony formation rate and cel cycle before and after transfection (P>0.05). And the alkaline phosphatase expressed positively. Infection for 48 hours at MOI 50 is optimal for transfecting dental pulp stem cel s with green fluorescent protein by a lentiviral vector, thereby providing reliable tracer method for the study of rat dental pulp stem cel s in vivo.

Objective To investigate the roles of sphingosine kinasel (SPK1) in apoptosis,invasiveness and multidrug resistance of human hepatocellular carcinoma cell line BEL-FU.Methods BEL-FU cells were infected with adenovirus carrying SPK1wT gene and SPK1siRNA (Ad-H1-SPK1) gene.Their effects on biological characteristics of BEL-FU cells were evaluated by MTT,cellular SPK enzyme activity assay,Transwell Migration Technology and Western-blot,respectively.Results AdSPK1wT significantly increased SPK activity but SPK1siRNA(Ad-H1-SPK1) decreased SPK activity.Over expression of SPK1 suppressed the apoptosis induced by DMS(Dimethyl sphingosine,DMS) and enhanced migration of BEL-FU cells.The cells infected with SPK1 siRNA( Ad-H1-SPK1)significantly increased the apoptosis induced by DMS and inhibited the migration of human hepatocellular carcinoma cells.The expression of multidrug resistance-related protein (MRP1) of cells infected with SPK1siRNA (Ad-H1-SPK1) was suppressed significantly compared with the control group,while the expression of MRP1 infected with Ad- SPK1wT was enhanced.Conclusion SPK1 activity is closely associated with apoptosis、migration and multidrug resistance of human hepatocellular carcinoma cells,therefore,it may serve as a new target for HCC treatment.

cryopreservation periodontal tissue. METHODS:Periodontal tissue was scraped from healthy human teeth and divided them into three equal parts:fresh group, harvesting periodontal ligament stem cel s from fresh tissue;5%dimethyl sulfoxide group, 5%dimethyl sulfoxide added into the cryopreservation system;10%dimethyl sulfoxide group, 10%dimethyl sulfoxide added into the cryopreservation system. One month later, periodontal ligament stem cel s were extracted from the cryopreserved periodontal ligament from the latter two groups. RESULTS AND CONCLUSION:In the time that cel s swam out of tissue mass and cel harvest amount, the 5%dimethyl sulfoxide group was inferior to the fresh group but better than the 10%dimethyl sulfoxide group (P<0.05). No differences were found among the three groups in the fol owing aspects (P>0.05):colony formation rate of passage 1 periodontal ligament stem cel s, cel survival rate, proliferation ability of passage 3 periodontal ligament stem cel s, cel growth curve and surface marker expression of periodontal ligament stem cel s. The results suggest that the 5%dimethyl sulfoxide added cryopreserved system for periodontal ligament tissue cannot only shorten the time of periodontal ligament stem cel s amplification in vitro, ensure cel harvest and maintain basic cel ular biological characteristics, but also reduce the total amount of the dimethyl sulfoxide and the direct damage to the cel s caused by repeatedly frozen cel s, thereby providing a more secure guarantee for the future implementation of clinical transplantation therapy. So, the 5%dimethyl sulfoxide added cryopreserved system may be the new selection for donor tissue storage.

Objective:To explore the latent classes of emotional behavior of children with mental disorders, and their relationship with parents' sense of parenting competence and psychological distress.Methods:A survey of 327 parents of children with mental disorders was conducted from September to December 2022 using the general information questionnaire, the sense of parenting competence scale, the Kessler psychological distress scale, and the strengths and difficulties questionnaire (parent version). Mplus 8.0 and SPSS 25.0 softwares were used for statistical analysis.The latent class analysis was used to identify subgroups of children with mental disorders based on their emotional behavior.Multinomial logistic regression was used to analyze the related factors.Results:Latent class analysis showed that the emotional behaviors of children with mental disorders were divided into 3 categories: emotion-conduct problem prominent group (38.53%(126/327)), simple emotional problem group (44.65%(146/327)), and emotion-peer interaction significant group (16.82%(55/327)). The differences among the 3 latent classes were statistically significant (all P<0.05) in terms of parents' parenting competence, satisfaction, and psychological distress scores.Compared with the emotion-conduct problem prominent group, the higher the parental parenting knowledge and parenting competence, the emotional behavior of children with mental disorders tended to be in the simple emotional problem group ( B=0.699, OR=2.011, 95% CI=1.046-3.868; B=0.088, OR=1.092, 95% CI=1.017-1.173). Compared with the " emotion-conduct problem prominent group" , the emotional behavior of children with mental disorders aged 13 to 18 years old tended to be in the " emotion-peer interaction significant group" ( B=1.982, OR=7.255, 95% CI=1.637-32.141). Conclusion:The emotional behavior of children with mental disorders is heterogeneous, and there are differences in sense of parenting competence and psychological distress of parents among different latent classes of children with mental disorders.

Objective:To investigate the effects of IL-28B in a mouse model of dextran sulfate sodium (DSS)-induced colitis and to analyze the possible mechanism.Methods:Thirty-five male C57BL/6 mice were randomly divided into the following groups with seven mice in each group: control group, DSS group and three IL-28B groups (1.25 μg, 2.5 μg and 5 μg). The mice in the DSS group and IL-28B groups were fed with 2.5% DSS solution and from day 3, the IL-28B groups were given intraperitoneal injection of corresponding IL-28B every day and the DSS group was treated with PBS. During the experiment, the disease activity index (DAI) was evaluated daily. On day 8, the mice were sacrificed and peripheral blood, spleen, mesenteric lymph node and colon samples were collected. The colon samples were observed, measured in length and stained with HE, and histopathological scores were calculated based on HE staining. Changes of immune cells in different samples were detected by flow cytometry. ELISA was used to detect the expression of IL-12, IL-10, IL-1β, IL-6, IL-4 and IL-13 in serum and colon tissues.Results:Compared with the DSS group, the IL-28B group (2.5 μg) had lower DAI scores [(9.40±1.67) vs (3.50±1.73), P<0.01], less shortening of the colon [(5.16±0.61) cm vs (6.91±0.60) cm, P<0.01] and significantly lower histopathological scores [(7.33±0.58) vs (4.33±0.58), P<0.01]. Moreover, compared with the DSS group, the IL-28B group (2.5 μg) showed decreased macrophages in the peripheral blood [(21.39±3.21)% vs (15.63±2.98)%, P<0.05] and spleen [(3.03±0.28)% vs (2.05±0.48)%, P<0.05], and significantly increased mean fluorescence intensity of M2 macrophages in the colon [(1 361.00±293.40) vs (2 074.00±87.61), P<0.05]. IL-12 expression in colon tissues and IL-1β expression in serum were reduced, and IL-10, IL-4 and IL-13 expression in colon tissues was significantly increased in the IL-28B group (2.5 μg) as compared with those in the DSS group [IL-12: (31.72±6.92) pg/mg vs (5.41±3.41) pg/mg; IL-1β: (48.01±16.13) pg/ml vs (12.27±6.26) pg/ml; IL-10: (184.70±46.82) pg/mg vs (444.30±157.80) pg/mg; IL-4: (2.23±0.27) pg/mg vs (3.64±0.80) pg/mg; IL-13: (11.79±0.99) pg/mg vs (22.59±1.92) pg/mg; all P<0.05]. Conclusions:IL-28B might alleviate the severity of acute enteritis in mice by increasing the secretion of IL-4 and IL-13, regulating macrophage differentiation and modulating the expression of inflammatory factors.

Pregnancy ; Female ; Humans ; Live Birth ; Calcium ; Oocytes ; Pregnancy, Multiple ; Pregnancy Rate ; Retrospective Studies ; Fertilization in Vitro ; Oocyte Retrieval

Objective:To explore the expression of serum miR-224-5p in PDAC and its significance for early clinical diagnosis.Methods:From August 2018 to April 2020, 40 patients with PDAC (11 patients with early PDAC, 29 patients with advanced PDAC), 21 patients with chronic pancreatitis and 40 healthy volunteer controls admitted in Qingdao Municipal Hospital were enrolled. The level of serum miR-224-5p in each group was detected by qRT-PCR method, and the correlation with clinicopathological parameters was analyzed. The receiver-operating characteristic (ROC) curve of miR-224-5p, CA19-9 and miR-224-5p combined with CA19-9 were drawn, and the sensitivity and specificity of the diagnosis were calculated.Results:The serum miR-224-5p levels in early PDAC group, middle and late PDAC group, chronic pancreatitis group and healthy control group were 3.21(2.01, 4.60), 4.70(3.50, 8.26), 1.72(1.02, 2.78) and 1.38(0.89, 2.11), respectively; and the level of serum miR-224-5p in the middle and late PDAC group was significantly higher than that in the early PDAC group, and that in the early PDAC group was significantly higher than that in the chronic pancreatitis group and the healthy control group, and all the differences were statistically significant ( P<0.05). The sensitivity of serum miR-224-5p combined with CA19-9, miR-224-5p, and CA19-9 in the diagnosis of overall PDAC was 95.0%, 85.0% and 67.5%, respectively; and the specificity was 70.0%, 82.5% and 87.5%, respectively. The sensitivity for early PDAC was 90.9%, 72.7% and 63.6%, and the specificity was 85.0%, 72.5% and 87.5%, respectively. MiR-224-5p combined with CA19-9 has the highest specificity in the diagnosis of PDAC. The level of serum miR-224-5p in patients with PDAC was correlated with TNM stage, lymph node metastasis and distant metastasis (all P values <0.05). Conclusions:The expression of serum miR-224-5p was significantly up-regulated in patients with early PDAC, and The level of serum miR-224-5p in patients with PDAC was correlated with TNM stage, lymph node metastasis and distant metastasis. The sensitity of serum miR-224-5p and miR-224-5p combined with CA19-9 for early PDAC diagnosis were superior to CA19-9 alone, which can be used as a potential sensitive biological marker for early screening of PDAC.