In growing children, growth plate cartilage has limited self-repair ability upon fracture injury always leading to limb growth arrest. Interestingly, one type of fracture injuries within the growth plate achieve amazing self-healing, however, the mechanism is unclear. Using this type of fracture mouse model, we discovered the activation of Hedgehog (Hh) signaling in the injured growth plate, which could activate chondrocytes in growth plate and promote cartilage repair. Primary cilia are the central transduction mediator of Hh signaling. Notably, ciliary Hh-Smo-Gli signaling pathways were enriched in the growth plate during development. Moreover, chondrocytes in resting and proliferating zone were dynamically ciliated during growth plate repair. Furthermore, conditional deletion of the ciliary core gene Ift140 in cartilage disrupted cilia-mediated Hh signaling in growth plate. More importantly, activating ciliary Hh signaling by Smoothened agonist (SAG) significantly accelerated growth plate repair after injury. In sum, primary cilia mediate Hh signaling induced the activation of stem/progenitor chondrocytes and growth plate repair after fracture injury.
Mice ; Animals ; Hedgehog Proteins/genetics* ; Receptors, G-Protein-Coupled/metabolism* ; Cilia/metabolism* ; Cartilage/metabolism* ; Regeneration

Objective:To explore the genetic characteristics and evolution of hantavirus carried by rodents in port area of Ningde in Fujian province in the summer of 2020.Methods:Rodents were captured in the port area of Ningde, the RNA was extracted from rodent lung tissues and detected by using specific kit. The positive samples were used for whole-genome sequencing of the virus. Bioinformatics software was used for the analysis on the similarity and genetic variation of the sequences.Results:A total of 112 rodents were captured, including 5 Rattus norvegicus and 2 Rattus flavipectus, the positive rate of hantavirus was 6.25% (7/112). By virus gene sequencing, two hantavirus complete genome sequences were obtained (named as FJ35 and FJ36, GenBank accession numbers: MW449188-MW449193). The genetic analysis results showed that the hantavirus detected in positive samples were SEOV and shared 99% nucleotide similarity with hantavirus strains LZSF21 and JX20140581 isolated from Shandong province. Phylogenetic analysis using the maximum likelihood method showed that the hantavirus detected in positive samples belonged to S3 subtype, sharing the same subtype with hantavirus strains Z37 from Zhejiang province, LZSF21 from Shandong province, and zy27 and Gongzhuling 415 from northeastern China. Compared with FJ372, the amino acid variation of N259S was observed at sites 251-264 of nucleoprotein, which might be related to antigenicity. Another variation of Q81R was observed in glycoprotein compared with SEOV 80-39 segment of coded amino acid of international reference strain, which might also cause the change in antigenicity. Conclusion:The high positive rate of hantavirus in rodents in the port area of Ningde- would increase the risk of natural human infection and epidemic in local area. The hantavirus positive rodents in this focus might be from an endemic area in Shandong. It is necessary to strengthen the imported rodent control in the port area of Ningde. The virus detected in 2 positive samples belonged to SEOV subtype Ⅲ and shared high homologies of nucleotides and amino acid sequences with the hantavirus strains in surrounding area. However, some slight variations occurred in glycoprotein and nucleoprotein amino acid sequences, which might cause changes in its antigeniity.

Objective:To search for the clinical indicators in differentiating Graves′ disease from subacute thyroiditis (SAT).Methods:Retrospective analysis was performed on thyroid function measurement of 265 cases of newly diagnosed Graves′ disease, 76 cases of SAT with thyrotoxicosis, 100 cases of non-toxic thyroid nodules, 105 cases of autoimmune thyroid diseases with normal thyroid function, and 151 cases of outpatients with normal thyroid function and without thyroid diseases.Results:Free triiodothyronine(FT 3)/free thyroxine(FT 4) ratio of Graves′ disease patients was significantly higher than that of SAT patients with thyrotoxicosis (0.65±0.29 vs 0.32±0.75, P<0.05). Receiver operating characteristic curve(ROC curve) analysis of FT 3/FT 4 ratio between Graves′ disease group and SAT group showed that FT 3/FT 4 ratio greater than 0.4 with a sensitivity of 98.11% and a specificity of 83.81% for diagnosis of Graves′ disease. Conclusion:FT 3/FT 4 ratio greater than 0.4 is helpful for differentiating Graves′ disease from subacute thyroiditis with thyrotoxicosis.

Objective To investigate the clinical effect of minimally invasive extraction of anterior tooth residual root after root separation.Methods A total of 400 patients receivinganterior tooth residual root extraction were collected in the clinic of oral and maxillofacial surgery department between January 2015 and December 2016.The patients were divided into a control group and a study group according to their sequence to see the doctor,with an odd for the study group and an even for the control group.In the study group,residual roots were separated mesiodistally by high speed turbine before using minimally invasive extraction tool;while in the control group residual roots were extracted only using minimally invasive extraction tool.The surgical duration,postoperative damage rate of the lip side plate,degree of pain and patient satisfaction in the two groups were analyzed.Results The surgical duration was shorter in the study group compared with the control group (P ＜ 0.05).The postoperative damage rate of the lip side plate and the degree of pain were lower,while patient satisfaction was higher in the study group than in the control group (P ＜ 0.05).Conclusions The postoperative damage rate of the lip side plate is significantly lower in minimally invasive extraction of anterior tooth residual root after root separation.Smaller trauma is conducive to the implant afterwards.Root separation in minimally invasive extraction of anterior tooth residual root is valuable for clinical application.

Objective S gene of hantavirus(HV) was expressed in insect cells by genetic engineering technology.The expression product of S gene was used as antigen to detect anti-HV specific antibody IgG in serum.Methods Gene encoding NP of the strain HV-Z10 was amplified by PCR and then its eukaryotic expression system rBAC-Z 10S-TN was constructed by using the routine genetic engineering method.SDS-PAGE was applied to measure the expression of rNP.Ion-exchange plus Ni-NTA-affinity chromatography was performed to purify the recombinant product.Indirect immuno-fluorescence assay (IFA) was used to determine the specific immune-reactivity of rNP.WB assay was established to detect the serum samples from 95 confirmed HFRS patients.Parameters related to the outcomes of detection were compared with the routine HV-IgG IFA method.Results rBAC-Z10S-TN was able to express rNP with high efficiency.The purified rNP only showed a single protein fragment in the gel after SDS-PAGE.HV IgG could efficiently recognize rNP and hybridize with the recombinant protein.97.67％ of the serum samples from the HFRS patients were positive confirmed by WB.Conclusions We successfully constructed a high efficient prokaryotic expression system of NP encoding gene from hantavirus strain HV-Z10.WB assay which was established in this study could be used as a new serological test for HFRS diagnosis,thanks to the simplicity,safety,sensitivity and specificity of this method.

Objective To cultivate human leukemia cells (HL-60) which were transiently transfected either a miR-126 mimic or inhibitor,and then to characterize the proliferation and apoptotic behavior of the transfected leukemia cells.Methods The leukemia cell line was developed and RT-PCR was performed to evaluate miR-126 expression levels.An instantaneous plasmid tmnsfection technique was used to transfect cells with either the miR-126 mimic or inhibitor.CCK-8,FCM,and clone formation tests were performed to analyze the proliferative and apoptotic behaviors of the leukemia cells.Results Proliferation was significantly decreased in cells transfected with the miR-126 mimic for 0,24,48,and 72 hours (P ＜ 0.05).Specifically,the G1,S,and G2 phases were significantly inhibited (P ＜ 0.05),the late and early apoptosis (UR+LR) rate increased (P ＜ 0.05),and the average rate of colony formation was also significantly decreased (P ＜ 0.05).Additionally,proliferation was significantly increased in cells transfected with the miR-126 inhibitor for 0,24,48,and 72 hours (P ＜ 0.05).Specifically,the G1,S,and G2 phases were increased (P ＜ 0.05),the UR + LR decreased significantly (P ＜ 0.05),and the average rate of colony formation was significantly increased (P ＜0.05).Conclusion In HL-60 cells,miR-126 can inhibit proliferation and promote apoptosis;thus,miR-126 may play an important role in the occurrence and development of leukemia as a tumor-suppressor miRNA.

Objective To investigate the effect and its mechanism of diazoxide on the blood-brain barrier (BBB) of rats after cerebral ischemia/reperfusion (I/R) injury.Methods Sixty Wistar rats were randomly divided into sham operation group,I/R group,and diazoxide pretreatment groups of low,middle,large dose (5,10,20 mg/kg).The I/R models of rats were performed to undergo middle cerebral artery embolism by thread.BBB permeability was estimated by Evans blue (EB) dyeing,transmission electron microscope (TEM) was used to observe the modification of interendothelial tight junction (TJ) of capillaries.The expression of aquaporin-4 (AQP4) in every rat brain tissues was detected by immunity histochemistry technique.Results (1) Compared to sham operation group,the permeability extent of EB were significantly increased by I/R,which was distinctly attenuated in middle and large dose of diazoxide pretreatment rats,while no obvious changes were found between I/R and low dose groups.(2) TEM showed that TJ of the brain tissue opened after I/R injury and no significant opening of TJ was observed in middle and large dose of diazoxide preconditioning groups.(3) Compared to sham operation group,the expression of AQP4 in the brain tissue of the I/R group was apparently increased (P ＜0.01).Compared to I/R group,the expression of AQP4 was apparently increased in middle and large dose pretreatment groups (P ＜ 0.01),and there were no obvious difference between low dose group and the I/R group.Conclusions Preconditioning of ischemia/reperfusion injury with diazoxide protects the blood-brain barrier,which may due to keep the TJ closed and decrease expression of AQP4 protein.

OBJECTIVE To study the molecular mechanism of cisplatin(DDP)by which HeLa cell growth and proliferation are inhibited. METHODS Cultured HeLa cells were treated with DDP 0.02-75 μmol · L-1 for 24 or 48 h. CCK-8 assay was used to determine the cell proliferation. The wound scratch assay was used to detect the cell migration and invasion. Flow cytometry was used to detect the cell cycle arresting. q-PCR was used to test the expression of metastasis suppressor gene 1 (MTSS1)mRNA. Western blot was used to determine protein levels of MTSS1,phosphorylated-extra?cellular signal-regulated kinase(p-ERK) and phosphorylated-serine-threonine kinase(p-AKT). RESULTS Following the treatment with DDP for 24 or 48 h,the proliferation of HeLa cells was inhibited significantly (P<0.05),the value of the half inhibitory concentration (IC50) of cells was 4.14 and 11.82 μmol · L-1. Migration and invasion activity of HeLa cells were reduced according to the wound scratch assay(P<0.05). Flow cytometry results showed that the cell cycle was arrested at S phase. q-PCR results showed that MTSS1 mRNA expression changed with DDP in a concentration-dependent manner (r24 h=-0.965,P<0.01;r48 h=-0.953,P<0.01). Western blot showed that the protein levels of MTSS1,p-ERK and p-AKT expression declined significantly with the increase in DDP concentrations(p-ERK:r24 h=-0.875,P<0.01;r48 h=-0.966,P<0.01. p-AKT:r24 h=-0.831,P<0.01;r48 h=-0.863,P<0.01. MTSS1:r24 h=-0.969,P<0.01;r48 h=-0.988,P<0.01). CONCLUSION DDP treatment inhibits HeLa growth and proliferation by interfering with the MTSS1 expression and disturbing the activation of ERK and AKT signaling pathways.

Objective To analyze labor duration in nulliparous women and discuss the change about curve of labor duration. Methods Two thousand, one hundred and forty nulliparous, full-term pregnant, singleton, cephalic presentation and vaginal delivery women who delivered at Peking University First Hospital were included. There were 1 300 cases between January 1, and December 31, 2009, while 840 cases between January 1, and May 31, 2013. A retrospective study was conducted. Data on maternal age, gestational age at delivery, body mass index at delivery, newborn weight, time of labor initiation, cervical dilatation and the amount of bleeding within the first 24 h after delivery were recorded. Data were compared by t test and χ2 test. The median time span corresponding to one-centimeter-increase in cervical dilatation was calculated. Results (1)Compared with data from 2009, the maternal age [(29.0±3.0) vs (29.6±2.8) years, t=4.77], incidence of postpartum hemorrhage [1.8%(24/1 300) vs 4.3%(36/840),χ2=11.17], proportion of induced labor [37.3%(485/1 300) vs 64.0%(538/840),χ2=146.23] and proportion of analgesia during labor [54.2%(705/1 300) vs 61.5% (517/840), χ2=11.15] were all higher in 2013 (all P<0.01). (2)The median (minimum–maximum) time span corresponding to a one-centimeter-increase in cervical dilatation was 3-4 cm which corresponded to 2.2 h(0.8-4.3 h), 4-5 cm which corresponded to 1.9 h(0.6-4.0 h), 5-6 cm which corresponded to 1.8 h(0.5-4.0 h), 6-7 cm which corresponded to 1.6 h(0.5-2.0 h), 7-8 cm which corresponded to 1.8 h(0.8-2.0 h), 8-9 cm which corresponded to 1.3 h(0.2-2.5 h), and 9-10 cm which corresponded to 0.6 h(0.1-2.5 h). The curve of labor duration showed a slow uptrend with time on the horizontal axis and cervical dilatation on the vertical axis. Conclusions The curve of labor duration exhibits a slow uptrend with neither an acceleration phase nor a deceleration phase. It is important to redefine the time span of labor duration in China for appropriate clinical treatment.

Objective To observe the effects of intensive training at different intensities on the expressions of semaphorin 3A ( Sema 3A) and its receptor neuropilin ( NP-1 ) and the cell apoptosis in cerebrum after cerebral ischemia-reperfusion in rats,and to investigate the possible mechanism of intensive training in recovery of motor function after cerebral ischemia-reperfusion in rats.Methods To establish animal model of cerebral ischemia-reperfusion in rats,the intraluminal thread method was applied to cause left middle cerebral artery occlusion (MCAO) for 2 h and before reperfusion.After cerebral ischemia-reperfusion model were established for 24 h,60 male model Wistar rats were randomly divided into training group 1 ( swimming for 5 min once a day),training group 2 ( swimming for 10 min once a day),training group 3 (swimming for 10 min twice a day) and control group (no training) ; another 15 rats assigned to the sham-operation group were subject to no MCAO and no training.Neurological function was evaluated by Garcia scores,and terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) was performed to dectect the cortical cell apoptosis.Expressions of neural growth inhibition factor Sema 3A and its receptor NP-1 were detected by immunohistochemistry.Results The neurological function in sham-operation group was normal.The differences of Garcia scores at different time points beween sham-operation group and control group were significant (P ＜ 0.01 ).Garcia scores in all training groups,were significantly higher than those in controls at the 7th and 14th d after swimming training ( P ＜ 0.01 ),especially in training group 3 the Garcia scores were ( 12.80 ± 0.45 ),( 15.20 ± 0.45 ),( 16.80 ± 0.45 ),respectively,at the 3rd,7th and 14th d after swimming training.The rates of positive cell of Sema 3A,NP-1 and TUNEL indexes in all training groups were lower than those in controls at the 3rd,7th and 14th d after swimming training (P ＜ 0.01 ),especially in the training group 3.At the 3rd,7th and 14th d after swimming training in training group 3,the rates of TUNEL indexes positive apoptosis cells were ( 29.43 ± 1.38 ) ％,( 22.30 ± 1.21 ) ％,( 17.58 ± 1.70) ％,respectively,the positive cell rates of Sema 3A were ( 19.64 ± 1.17) ％,(9.73 ± 3.83)％,(8.24 ± 0.87)％,respectively,the positive cell rates of NP-1 were ( 33.95 ± 6.86) ％,( 27.95 ± 1.29 ) ％,( 18.90 ± 1.44 ) ％,respectively,the reduction of positive cells expressions in training group 3 was significantly more obvious compared with other training groups (P ＜ 0.01 or 0.05).Conclusions Rehabilitation training can reduce the expression of positive cell of Sema 3A,NP-1 and TUNEL indexes in rats after cerebral ischemia-reperfusion and can improve motor function recovery and facilitate neural plasticity.The more intensive the training,the better the effects.