BACKGROUND: The addition of growth factiors is commonly applied to improve the osteogenic differentiation of stem cells. However, for human pluripotent stem cells (hPSCs), their complex differentiation processes result in the unknown effect at different stages. In this study, we focused on the widely used bone forming peptide-1 (BFP-1) and investigated the effect and mechanisms of its addition on the osteogenic induction of hPSCs as a function of the supplementation period. METHODS: Monolayer-cultured hPSCs were cultured in osteogenic induction medium for 28 days, and the effect of BFP-1 peptide addition at varying weeks was examined. After differentiation for varying days (0, 7, 14, 21 and 28), the differentiation efficiency was determined by RT–PCR, flow cytometry, immunofluorescence, and alizarin red staining assays. Moreover, the expression of marker genes related to germ layers and epithelial-mesenchymal transition (EMT) was investigated at day 7. RESULTS: Peptide treatment during the first week promoted the generation of mesoderm cells and mesenchymal-like cells from hiPSCs. Then, the upregulated expression of osteogenesis marker genes/proteins was detected in both hESCs and hiPSCs during subsequent inductions with BFP-1 peptide treatment. Fortunately, further experimental design confirmed that treating the BFP-1 peptide during 7–21 days showed even better performance for hESCs but was ineffective for hiPSCs. CONCLUSION The differentiation efficiency of cells could be improved by determining the optimal treatment period.Our study has great value in maximizing the differentiation of hPSCs by adding osteogenesis peptides based on the revealed mechanisms and promoting the application of hPSCs in bone tissue regeneration.

Objective:To investigate the mid-term clinical outcomes of arthroscopic vertical mattress suturing for shoulder recurrent anterior dislocation combined with joint laxity.Methods:A retrospective case series study was performed on the clinical data of 11 patients with recurrent anterior shoulder dislocation combined with joint laxity admitted to the First Affiliated Hospital, Army Medical University from January 2018 to September 2021. The patients included 10 males and 1 female, aged 18-38 years [(22.8±5.5)years]. All the patients received treatment with arthroscopic vertical mattress suturing. The Oxford shoulder instability score, Rowe shoulder instability score, and simple shoulder test (SST) score were compared before operation, at 6 months after operation and at the final follow-up. The degree of joint capsule laxity and length of capsular redundancy (evaluated by MRI) were compared before operation and at the final follow-up. The results of the supine apprehension test, re-dislocation and postoperative complications such as iatrogenic vascular and nerve injuries were observed at the final follow-up. Also, the correlation between the radiological changes in the joint capsule and the shoulder function was analyzed by Spearman correlation coefficient.Results:All the patients were followed up for 20-64 months [(40.7±18.6)months]. Before operation, at 6 months after surgery and at the final follow-up, the values of Oxford shoulder instability score were (41.2±4.7)points, (49.5±3.0)points and (57.6±3.0)points; the values of Rowe shoulder instability score were (28.6±9.5)points, (77.7±7.2)points and (94.1±10.9)points; and the values of SST score were (7.6±1.3)points, (9.8±1.0)points and (11.6±0.9)points, respectively. The Oxford shoulder instability score, Rowe shoulder instability score and SST at 6 months after operation and at the final follow-up were significantly better than those before operation, and those at the final follow-up were significantly better than those at 6 months after operation (all P<0.05). The MRI showed that the degree of joint capsular laxity and length of capsular redundancy were 1.5±0.2 and (19.7±2.5)mm before operation and were 1.3±0.2 and (12.9±3.7)mm at the final follow-up, respectively ( P<0.05 or 0.01). The supine apprehension test was negative at the final follow-up, with no re-dislocation or postoperative complications such as iatrogenic vascular or nerve injuries. Correlation analysis showed a negative correlation between the degree of joint capsular laxity and the Oxford shoulder instability score ( r=-0.62, P<0.05) and that of the length of capsular redundancy with the Oxford shoulder instability score ( r=-0.80, P<0.01), the Rowe shoulder stability score ( r=-0.73, P<0.01) and the SST score ( r=-0.75, P<0.01). Conclusions:Arthroscopic vertical mattress suturing has good mid-term clinical outcome for recurrent shoulder anterior dislocation combined with joint laxity, improving the shoulder function and reducing complications, wihch is associated with decreased joint capsule laxity and length of capsular redundancy.

Objective:To investigate the effect of paeoniflorin on toll-like receptor 4(TLR4)/nuclear transcription factor(NF-κB) signaling pathway of streptozotocin combined with ovariectomized mice, and to explore whether it can improve the cognitive impairment of ovariectomized diabetic mice.Methods:Ninety female C57BL/6J mice were divided into SHAM group, ovariectomy group, diabetes group(intraperitoneal injection of STZ 50 mg·kg -1·d -1 for 5 consecutive days), dual model group(DM modeling and OVX operation), paeoniflorin low-dose intervention group(OVX+ STZ+ L-PF 50 mg·kg -1·d -1), paeoniflorin high-dose intervention group(OVX+ STZ+ H-PF 100 mg·kg -1·d -1; all groups n=15). After 8 weeks of paeoniflorin intervention, their cognitive function was tested by behavioral experiments(Morris water maze and Y maze). The pathological changes of hippocampal tissue were observed by HE and Nissl staining. The mRNA expressions of TLR4, tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β) and interleukin-6(IL-6) in hippocampal tissues were detected by real-time fluorescence quantitative PCR. The expression of TLR4, NF-κB P65, TNF-α, IL-6, IL-1β, β-amyloid protein(Aβ), tau proteins, and p-tau proteins were detected by Western blot. Results:Compared with SHAM group, the learning and memory ability of ovariectomy group, diabetes group and dual model group decreased, hippocampal cells were damaged, and the expression of related gene mRNA and protein were increased, especially in dual model group; Compared with dual model group, paeoniflorin intervention could delayed the learning and memory impairment, improve cognitive function, reduce the degree of hippocampal injury, and decrease the expression levels of related gene mRNA and protein, The above changes were the most pronounced at paeoniflorin high-dose intervention group.Conclusion:Paeoniflorin improves cognitive dysfunction in ovariectomized diabetic mice by inhibiting TLR4/NF-κB signaling pathway.

Background/Aims: Existing hepatocellular carcinoma (HCC) prediction models are derived mainly from pretreatment or early on-treatment parameters. We reassessed the dynamic changes in the performance of 17 HCC models in patients with chronic hepatitis B (CHB) during long-term antiviral therapy (AVT). Methods: Among 987 CHB patients administered long-term entecavir therapy, 660 patients had 8 years of follow-up data. Model scores were calculated using on-treatment values at 2.5, 3, 3.5, 4, 4.5, and 5 years of AVT to predict threeyear HCC occurrence. Model performance was assessed with the area under the receiver operating curve (AUROC). The original model cutoffs to distinguish different levels of HCC risk were evaluated by the log-rank test. Results: The AUROCs of the 17 HCC models varied from 0.51 to 0.78 when using on-treatment scores from years 2.5 to 5. Models with a cirrhosis variable showed numerically higher AUROCs (pooled at 0.65–0.73 for treated, untreated, or mixed treatment models) than models without (treated or mixed models: 0.61–0.68; untreated models: 0.51–0.59). Stratification into low, intermediate, and high-risk levels using the original cutoff values could no longer reflect the true HCC incidence using scores after 3.5 years of AVT for models without cirrhosis and after 4 years of AVT for models with cirrhosis. Conclusions The performance of existing HCC prediction models, especially models without the cirrhosis variable, decreased in CHB patients on long-term AVT. The optimization of existing models or the development of novel models for better HCC prediction during long-term AVT is warranted.

Maili moxibustion can alleviate cancer pain, reduce bone marrow suppression, alleviate gastrointestinal reaction of chemotherapeutic drugs, alleviate cancer-related fatigue, inhibit neurotoxic reaction, improve quality of life and prolong patients' survival. It plays therapeutic effects by regulating immunity, inhibiting tumor cell proliferation and regulating tumor microenvironment. The researches of Maili moxibustion for tumor focus on reducing the toxic or side effects of radiotherapy or chemotherapy. In the future, we should continue to study the combination of Maili moxibustion and other therapies on the treatment of tumor.

Objective: The cross-sectional study examined associations between gross motor competence and physical fitness in 3-5 years old children. Methods: A convenient sampling method was used to select 201 children aged 3-5 years, to assess gross motor competence, each child accomplished the Test of Gross Motor Development-3(TGMD-3). The level of physical fitness was measured by National Physical Fitness Test, comprising eight different tasks including height, weight, flexibility(sit-and-reach test), balance (walking on a balance beam), explosive force(Standing long jump), strength(Tennis ball throwing), agility(10-shuttle-run test), strength and coordination(Continuous foot jump). Results: With the increase of age, the scores of locomotor skills，object control skills, gross motor competence, height, weight, walking on a balance beam, continuous foot jump, tennis ball throwing and standing long jump were better(P<0.01). Significant differences were not found in the gross motor competence, locomotor skills, 10-shuttle-run test, continuous foot jump, standing long jump and walking on a balance beam(P>0.05). In the object control skills and tennis ball throwing, the boys showed a greater performance(P<0.01). In the sit-and-reach test, girls showed a greater performance(P<0.01). The results revealed a significant relationship between gross motor competence and the following items: 10-shuttle-run test, continuous foot jump, standing long jump, walking on a balance beam, tennis ball throwing(P<0.05). There were no associations among gross motor competence, BMI(r=0.02), sit-and-reach test(r=0.07). There were no associations between gross motor competence and physical fitness in 3-5 years old(r=-0.13-0.21), except for 5 years old children’s object control skills and continuous foot jump(r=0.42). Sex, age, height , BMI (R2=0.05,F=2.62, P>0.05) as well as gross motor competence(△R2=0.04, F=2.00, P>0.05) did not predict physical fitness. Conclusion The results indicated a relationship between motor competence and physical fitness in 3-5 years old. In order to increase the level of physical fitness, the educators should pay attention to the development of children’s gross motor.

Objective To explore the effect and mechanism of Kuntai Capsule (KC) on premature ovarian failure in rats.Methods Totally 40 SD female rats were randomly divided into four groups:control group,model group,Kuntai Capsule (KC) group and conjugated estrogens tablets (CET) group.The premature ovarian failure model in rats was made by ig administration of 75 mg/kg tripterygium.After the model was established,rats were continually treated with 0.6 and 0.625 g/kg of KC and CET respectively by ig administration for 36 d.HE staining was used to observe the morphology of ovary and count the number of follicle.The enzyme-linked immunosorbent assay (Elisa) method was used for the detection of follicle stimulating hormone (FSH),luteinizing hormone (LH),serum estradiol (E2),and anti-Mueller tube hormone (AMH) level.The real-time fluorescence quantitative PCR (qRT-PCR) method was used to detect the mRNA expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF).Results Compared with control group,the number of primordial follicles,preantral follicles and antral follicles were significantly reduced,the number of atretic follicles was increased (P ＜ 0.05),E2 and AMH significantly decreased (P ＜ 0.05),FSH and LH levels increased significantly (P ＜ 0.05),and the levels of VEGF and bFGF mRNA in model group were significantly decreased (P ＜ 0.05).Compared with model group,the number of primordial follicles,preantral follicles,and antral follicles were significantly increased,the number of atretic follicles was significantly reduced (P ＜ 0.05),E2 and AMH significantly increased (P ＜ 0.05),FSH and LH levels significantly decreased (P ＜ 0.05),and the levels of VEGF and bFGF mRNA in KC group were significantly increased (P ＜ 0.05).Conclusion By regulating the level of hormone and up-regulating the expression of VEGF and bFGF,KC can repair the damaged ovarian tissue and promote the growth and development of the follicle,so as to inhibit the premature exhaustion of mRNA.

Aim To observate the effect of chemokine receptor(CXCR4) gene transfection on biological behavior of bone marrow mesenchymal stem cells in vitro.Methods Firstly, bone marrow mesenchymal stem cells were divided into three groups:GFP(transfected GFP into MSCs), CXCR4+(transfected CXCR4+ into MSCs) and CXCR4-(transfected CXCR4-into MSCs) group.Then, their capacity of proliferation, differentiation and migration ability (in vitro) was assessed with immunofluorescence cytochemistry method, flow cytometry assay and Transwell cell chemotaxis test.Results The high or low expression of CXCR4 had no effect on their ability of proliferation and differentiation into lung tissue.Compared with GFP group, however, CXCR4+-MSCs group significantly increased the number of migrating cells, while CXCR4——MSCs group showed no change in the number of migrating cells.Conclusions The proliferation and differentiation capacities are not affected by the high or low expression of CXCR4.The high expression of CXCR4 can significantly enhance the migration ability of MSCs to inflammatory lesions, and the low one has no effect on the migration of the cells.After the transplantation of MSCs, CXCR4′s high expression will access to the lesion area to participate in tissue repairing rapidly and largely, significantly enhancing the therapeutic efficacy.

Objective To investigate the abnormal functional connectivity (FC) between the substantia nigra (SN) and the brain motor area in rapid eye movement sleep behavior disorder (RBD) by Unified Parkinson′s Disease Rating Scale (UPDRS), Hoehn-Yahr Scale and resting state functional magnetic resonance imaging (rfMRI).Methods A total of 34 subjects (14 with RBD (RBD group), 12 with Parkinson′s disease (PD group), and 8 age, sex-matched normal controls (control group)) recruited in the Department of Neurology, Henan Provincial People′s Hospital from 2014 to 2015 were enrolled in this study according to international diagnosis criteria.All subjects were examined by UPDRS, Hoehn-Yahr Scale and rfMRI.Results UPDRS scores and Hoehn-Yahr staging were 0.00 (0.00, 3.75) and 0.00 (0.00, 0.50) respectively in the RBD group, which were significantly different from that in the PD group (30.5 (18.75, 33.00) and 1.75 (1.50, 2.50), respectively;Z=-3.782, P<0.05 and Z=-2.963, P<0.05), whereas not significantly different from that in the control group (0.00 (0.00, 0.00),0.00 (0.00, 0.00), respectively;Z=1.16, P>0.05 and Z=1.10, P>0.05).The FC analysis showed alterations from the right SN to bilateral cerebellum respectively among the three groups (Fright cerebellum=12.975, Fleft cerebellum=6.144;P<0.05);furthermore, there were significant increases in the RBD group compared with the PD group (t=-4.602,-5.080, P<0.005).The FC analysis showed reduced correlation from the right SN to the left prefrontal cortex in the RBD and the PD groups compared with the control group (t=-4.899,-3.500, P<0.005), but there was no statistically significant difference between the RBD and the PD groups (t=2.035, P>0.05).Conclusions There was abnormal FC from the SN to motor areas in RBD patients, and partial alterations were similar as PD patients.rfMRI provided an evidence that RBD might be presymptom of PD.

Objective A set of intracranial pressure and intracranial temperature monitor was developed. Moreover, it was verified to be effective in the monitoring of intracranial parameters by designed experiments.Methods The intracranial pressure and intracranial temperature monitor was tested in the water bath comparing with the Codman intracranial pressure monitor and mercury thermometers. As wel, the monitor was applied in the monitoring of rat brain edema in vivo.Results The maximum error is less than 266.64 Pa in the intracranial pressure measurement compared to the Codman intracranial pressure monitor, and the maximum error is less than 0.3oC in the temperature measurement according to mercury thermometers. Furthermore, the monitor could real-time obtain the intracranial pressure and intracranial temperature in the brain edema in vivo.Conclusion The intracranial pressure and intracranial temperature monitor realizes the real-time in vivo monitoring of intracranial pressure and intracranial temperature. The measurement accuracy meets the acquirement of doctors. The instrument has potential for clinical use.