Objective: To study the effect of ultrasonic guidance in radial artery catheterization. Methods: From January 2017 to January 2018, 100 patients who received surgical treatment in Taizhou Central Hospital of Zhejiang Province and needed puncture and catheterization anesthesia during operation were randomly divided into control group (n=50) and observation group (n=50) according to digital table method.The control group adopted the traditional touch pulse method, while the observation group was guided by ultrasound.The first puncture success rate, the second puncture success rate, puncture time, puncture times, the vital sign index, incidence of adverse events, pain score, comfort score were compared between the two groups. Results: The first puncture success rate (80%) and the second puncture success rate (100%) of the observation group were higher than 62% and 88% of the control group (χ²=3.934, 6.383, all P<0.05). The single puncture time [(59.85±8.31)s] and total puncture time [(107.14±35.37)s] of the observation group were shorter than those of the control group (t=5.766, 6.398, all P<0.05), and the puncture times [(1.27±0.54) times] of the observation group was less than that of the control group (t=5.689, P<0.05). After puncture, the heart rate [(73.79±2.46)times/min] and average arterial pressure [(96.75±6.76)mmHg] of the observation group were all lower than those of the control group (t=5.058, 4.635, all P<0.05). The total incidence of adverse events such as subcutaneous hematoma and arterial wall penetration in the observation group (4%) was lower than that in the control group (18%) (χ²=5.005, P<0.05). The pain scores at puncture and after puncture of the observation group [(1.53±0.71)points, (1.28±0.60)points] were lower than those of the control group (t=5.545, 6.233, all P<0.05). The comfort scores at puncture and after puncture of the observation group [(91.25±9.64)points, (97.31±11.37)points] were higher than those of the control group (t=6.147, 6.226, all P<0.05). Conclusion Ultrasonic guidance can effectively improve the puncture efficiency, reduce the fluctuation of vital signs, pain, reduce the adverse events during puncture, and make the patients feel more comfortable.

OBJECTIVETo establish a human embryonic stem cell line with stably β-catenin gene silencing by lentivirus-mediated shRNA interference.

METHODSPLKO.1-puro-β-catenin vector, a lentivirus plasmid expressing β-catenin shRNA, was packaged into 293FT cells. Human embryonic stem cells were infected with the lentivirus and the cell clones stably expressing β-catenin shRNA were selected by puromycin, with the uninfected cells and cells infected with the empty vector as the control. Real-time RT-PCR was used to evaluate the efficiency of β-catenin knocked down; β-catenin and OCT4 protein expression in the infected cells was examined using immunofluorescence assay.

RESULTSInfection with β-catenin-specific shRNA lentivirus resulted in stable interference of β-catenin expression in human embryonic stem cells, which showed a β-catenin mRNA expression of only 1% of that in the uninfected cells. Infection with the empty vector produced no obvious effect on β-catenin expression compared to the uninfected cells. In the cells infected with β-catenin shRNA lentivirus, β-catenin protein expression was almost undetectable in immunofluorescence assay, while OCT4 was still expressed after the interference.

CONCLUSIONLentiviral vector-delivered shRNA results in effective and stable β-catenin gene silencing in human embryonic stem cells.

Cell Line ; Embryonic Stem Cells ; cytology ; metabolism ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Plasmids ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; Wnt Signaling Pathway ; beta Catenin ; genetics ; metabolism

Objective To explore the pathogenesis of steroid-induced osteonecrosis of the femoral head(SONFH)and the mechanism of Traditional Chinese Medicine(TCM)-based prescription for"Bringing Blood Downward"targeting SONFH through the OPG/RANKL/RANK signaling pathway.Methods Fifty male SD rats were randomly divided into model group and control group.The rats in the model group were treated with LPS and methylprednisolone to build a SONFH model.After 6 weeks,two rats were randomly selected respectively from the two groups,and the SONFH model was confirmed by histomorphological examination under light microscopy.The rats in a low,medium and high dose treatment groups(group L,M,and H)were intragastrically administrated with the Prescription for"Bringing Blood Downward"at concentrations of 2.5/5/10 g/kg.While the rats in the model group and control group were intragastrically administrated with normal saline at a concentration of 5 g/(kg·d).The pathological changes and expression levels of OPG,TRAF-6,and RANKL in each group were examined 4 weeks later.Results Compared with the model group,chondrocytes and cell matrix of groups L,M and H were less reduced,osteoclastic absorption and trabecular structure were less destroyed and the cells were less adipose.The adipocyte density in the control group and each treatment group was significantly lower than that in the model group(P<0.01).In the control group,the bone marrow cavity area was smaller than that in the model group(P<0.05),but there was no significant difference between each dose treatment groups and the model group(P>0.05).The expression levels of OPG,RANKL,TRAF-6 and OPG/RANKL ratio in the control group and groups L,M and H were significantly lower than those in the control group(P<0.01).Conclusion The TCM-based prescription for"Bringing Blood Downward"works to treat SONFH byregulating the activity of osteoclasts and osteogenic differen-tiation process of osteoblasts,improving the microcirculation environment of the femoral head,and inducing the formation of trabecular bone through the OPG/RANKL/RANK signaling pathway.It functions with its multitargetsin diverse methods.

ObjectiveTo explore the mechanism of Danggui Niantongtang （DGNTT） against adjuvant-induced arthritis （AA） in rats with wind-dampness-heat arthralgia （FSR） based on the variation of intestinal flora. MethodA total of 60 SD rats were randomized into normal （control） group， FSR group， low-， medium-， and high-dose DGNTT （5.67， 11.34， 22.68 g·kg^-1） groups， and methotrexate （MTX） group （1.35 mg·kg^-1）， with 10 rats in each group. The rats， except the control group， were injected with Mtb adjuvant and then exposed to artificial climatic chamber （hot and humid with wind） for 64 h for modeling. The rats were treated with water， DGNTT or MTX for 28 days from the day of injection. Arthritis index （AI） of rats was measured and paw volume was determined with a volume meter. The morphology of synovial tissues of the knees was observed based on hematoxylin-eosin （HE） staining and the changes of intestinal flora were analyzed based on 16S rRNA sequencing. ResultDGNTT can alleviate the hyperplasia of synovial tissue and inflammation of AA rats with FSR and inhibit the formation of pannus. The results of 16S rRNA sequencing showed that the relative abundance of Firmicutes， Lactobacillus， Prevotella 9， and Alloprevotella decreased （P<0.05， P<0.01） and the relative abundance of Bacteroidetes and Bacteroides increased （P<0.01） in FSR group compared those in the control group. Compared with the FSR group， all DGNTT groups and MTX group had high relative abundance of Lactobacillus （P<0.05， P<0.01） and low relative abundance of Bacteroidetes （P<0.01） and medium-dose and high-dose DGNTT groups and MTX group showed high abundance of Firmicutes， Prevotella 9， and Alloprevotella and low abundance of Bacteroides （P<0.05， P<0.01）. Spearman's correlation analysis suggested that the abundance of Bacteroides and Helicobacter was in positive correlation with AI （P<0.05）， while the abundance of Prevotella 9 and Candidatus Saccharimonas was in negative correlation with AI （P<0.01， P<0.05）. There was a negative correlation between the abundance of Prevotella 9 and paw volume （P<0.01）， and the abundance of Ruminococcaceae NK4A214 group， Christensenellaceae R-7 group， and Bacteroides was in negative correlation with spleen index （P<0.05）. The abundance of Prevotella 9 was in negative correlation with spleen index （P<0.01）. ConclusionDGNTT is effective for arthritis with FSR， as it can regulate the composition of intestinal flora in AA rats by increasing the abundance of probiotics and inhibiting the growth of pathogenic bacteria. The mechanism is the likelihood that it improves intestinal immune metabolism to ensure intestinal homeostasis.