Objective Getting the robust exogenous gene expression vector under the control of porcine insulin promoter, and to lay the foundation for pancreaticβ-cells specific transgene expressing pigs.Method Using porcine insu-lin promoter ( PIP, 1500 bp of the 5′UTR from the porcine INS gene including the first exon and the first intron) to con-struct expression vector, the HindIII restriction site which connected the sequences of PIP and EGFP was designed before ATG, named PIP-HindIII-EGFP.Considering that the different location of restriction site may affect the expression efficien-cy of the transgene, we optimized the expression vector.Firstly the HindIII restriction site was deleted to realize the seam-less connection of PIP and EGFP,the vector was named PIP-EGFP.Also we mutated the 3′intron splicing acceptor site( SA) of the first intron into HindIII restriction site, named as PIP-SA( M)-EGFP.Three different EGFP expression vectors were respectively transfected MIN-6 mouse pancreatic β-cells, pig ear fibroblasts and kidney cells.The transfected cells were cultured for 48 h and harvested for RT-PCR, flow cytometry and Western blot analysis, to analyze and compare the expres-sion efficiency of vectors.Results After transfection,green fluorescence was observed only in MIN-6 mouse pancreaticβ-cells.RT-PCR analysis and product sequencing showed that the three expression vectors did have different stability with in-tron splicing.The PIP-HindIII-EGFP construct and PIP-EGFP vector produced two kinds of mRNA with the first intron spliced and no spliced, indicating the instability of intron splicing.Mutation of the PIP splice site would cause the first in-tron not spliced, while flow cytometry and Western blot displayed that the mutation induced a most efficient expression of the downstream gene.Conclusions A robust and specific β-cells expression vector has been successfully generated by mutating the intron splicing acceptor site of the porcine insulin promoter.It provides the foundation for preparation of pigs with pancreaticβ-cells specifically expressing the transgene.

Objective The objective of this study was to investigate the miR-345 expression level in non-small cell lung cancer(NSCLC)tissue and its potential clinical significance.Methods Real-time PCR was conducted to evaluate the expression of miR-345 in NSCLC tissues in 98 samples and cell lines.Then the association between tissue miR-345 expression level and clinical outcomes was further analyzed.Results The expression level of miR-345 was significantly decreased in NSCLC tissues and cell lines when compared to the controls(P<0.05).miR-345 expression level in NSCLS tissues was associated with various clinicopathological parameters including LN metastasis(P<0.001),distant metastasis(P=0.028),TNM stage(P=0.004)and grade(P=0.011).In addition,the NSCLC patients in the low miR-345 expression group showed significantly shorter overall survival time than that in the high miR-345expression group(P=0.021).Multivariate analysis showed that the expression of miR-345 was an independent risk factor for NSCLC(HR=3.897,95% CI:2.263~10.440;P=0.012).Conclusion The expression level of miR-345 was decreased in NSCLC tissues as well as cell lines compared with normal controls.Low miR-345 expression in tissues was associated with progression and poor prognosis of NSCLC.These results indicate that miR-345 may be a novel prognostic marker in NSCLC.

OBJECTIVETo explore the relationship between the needling sensation of catgut embedding therapy and the depth of embedded catgut so as to improve the safety of the needle insertion and catgut implantation of the therapy.

METHODSTwenty healthy adults were selected. Under the ultrasound, the structure of the cervical Jiaji (EX-B 2) was observed. In the ultrasound guidance, the catgut was embedded. The two-dimensional imaging method was adopted to observe the anatomic structure and the procedure of needle insertion at the cervical Jiaji (EX-B 2). The high-frequency ultrasound was used to collect the images at Jiaji (EX-B 2) of C5 and determine the depths from the skin surface to the different layers of the point. Additionally, the visual analogue scale (VAS) was adopted to score the needling sensations when the needle inserted at different layers. The persistent sensation duration in the local area was followed continuously.

RESULTSUnder the ultrasound, the anatomic structure and tissue layers of cervical Jiaji (EX-B 2) were displayed clearly. The difference was significant in the average depth from the skin surface to the subcutaneous tissue, trapezius, splenius capitis, semispinalis capitis, semipinalis cervicis, multifidus and vertebral arch between the males and females (all P<0. 01). During the needle insertion, the sensations were apparently different when the implantation went to different layers. The qi arrival presented when the catgut was embedded to the trapezius, splenius capitis, semispinalis capitis, semipinalis cervicis and multifidus. But the distending pain was the most significant when in the myofascial. Commonly, the embedded catgut 2. 5 cm in length may be implanted deeply to the multifidus and the local needling sensation lasted averagely for (72. 0 ± 10. 2) h. Conclusion Under the ultrasound guidance, the depth of embedded catgut is clearly displayed at cervical Jiaji (EX-B 2). The needle insertion and the implanted material are visible, and the relationship between qi arrival and the layer of needle insertion is determined. The accuracy and safety of minimally invasive catgut embedding therapy is improved in the treatment of cervical spondylosis.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Catgut ; utilization ; Female ; Humans ; Male ; Middle Aged ; Neck Muscles ; anatomy & histology ; diagnostic imaging ; Qi ; Sensation ; Spondylosis ; diagnostic imaging ; therapy ; Ultrasonography

Objective To investigate the antibacterial properties and curative effects of a nanosilver-epidermal growth factor (NanoAg-EGF) sustained-release carrier.Methods First,in accordance with former method,sustained-release carrier of NanoAgEGF was prepared,with the design of the NanoAg-EGF group,NanoAg-alone group,EGF-alone group,control groups of normal saline-alone,and benzylpenicillin sodium.Contrasting detection on antibacterial properties of five kinds of common microbes (staphylococcus aureus,escherichia coli,pseudomonas aeruginosa,candida albicans,and streptococcus pneumonia) in each group was used to confirm their antibacterial properties.Then,wound models of diabetic rats were randomly divided into five groups,with the wound treatments by sustained-release cartier of NanoAg-EGF,combined application of NanoAg + EGF,EGF-alone,NanoAg-alone,and normal saline-alone control group,respectively.The observation on the healing situation of each group in different times was used to ensure the curative effect.Results In the antibacterial experiments,it was found that the NanoAg-EGF and NanoAg had strong antibacterial properties.Their antibacterial properties had no significant difference (P ＞ 0.05).The control group,penicillin sodium only had relatively weak antibacterial properties on staphylococcus aureus and streptococcus pneumoniae,with obviously weaker function compared to the NanoAg-EGF and NanoAg groups (P ＜ 0.05).The control groups,EGF-alone and normal saline-alone had no inhibitor function on each kind of pathogenic microorganisms.In the wound healing experiments on diabetic rats,it was found that NanoAgEGF shortened significantly the wound healing time of diabetic rats (P ＜ 0.05).On the 3rd day,the healing rate of each group had no difference (P ＞ 0.05).On the 7th day and 13th day,the healing rates of NanoAg-EGF [(61.71 ± 8.78)％ and (100 ± 2.60) ％] were significantly higher than the other four groups (P ＜ 0.05).Conclusions NanoAg-EGF not only has good antibacterial properties but also can promote rapidly the wound healing of diabetes.

BACKGROUND: Hypoxic training in combined with exercise training can increase the hypoxia levels of the body to mobilize the potentials of the body, resist to and actively adapt physiological responses resulted from hypoxia, so as to improve athletic performance.OBJECTIVE: To summarize the influence of hypoxic training on skeletal muscle structure and function, to provide a theoretical basis and guidance for hypoxic training.METHODS: Using "hypoxic training, skeletal muscle, function, structure" in English as the search terms, PubMed database was ratrieved from 1990 to March 2009; Using "hypoxic training, skeletal muscle, function and structure" in Chinese as the search terms,CNKI database was searched from 2000 to May 2009. Literatures are limited to English and Chinese languages. The content related to the effect of hypoxic training on skeletal muscle structure and function was selected, and repetitive researches were eliminated.RESULTS AND CONCLUSION: Totally 144 literatures were obtained from computer screening, and then analyzed according to the inclusion and exclusion criteria. Hypoxic training could avoid the disadvantages caused by conventional altitude training, the hypoxia environment is regulated according to the body's ability to adapt hypoxia, the normal training is ensured, so that skeletal muscle obtains beneficial physiological adaptation. Compared with pure hypoxia stimulation, different forms of hypoxia and different training methods may integrate into different modes of hypoxic training, which makes skeletal muscle function and structure more complex. The combination form of hypoxia and training is the main factor of the effect of hypoxia training on skeletal muscle function and structure. Different objects of experiments and test methods are likely to interfere with the conclusions, which also affect the mechanism of the role of hypoxic training on skeletal muscle.

Objective:To evaluate facial soft tissue 3-deminsion changes of skeletal Class Ⅲmalocclu-sion patients after orthognathic surgery using structure light scanning technique .Methods:Eight patients [3 males and 5 females, aged ( 27.08 ±4.42 ) years ] with Class Ⅲ dentoskeletal relationship who underwent a bimaxillary orthognathic surgical procedure involving advancement of the maxilla by Le FortⅠosteotomy and mandibular setback by bilateral sagittal split ramus osteotomy (BSSO) and genioplasty to correct deformity were included .3D facial images were obtained by structure light scanner for all the patients 2 weeks preoperatively and 6 months postoperatively .The facial soft tissue changes were evalua-ted in 3-dimension.The linear distances and angulation changes for facial soft tissue landmarks were ana-lyzed.The soft tissue volumetric changes were assessed too .Results: There were significant differences in the sagittal and vertical changes of soft tissue landmarks .The greatest amount of soft tissue change was close to lips.There were more volumetric changes in the chin than in the maxilla , and fewer in the forehead .Conclusion: After biomaxillary surgery , there were significant facial soft tissue differences mainly in the sagittal and vertical dimension for skeletal Class Ⅲ patients .The structure light 3 D scan-ning technique can be accurately used to estimate the soft tissue changes in patients who undergo orthog-nathic surgery .

Objective To explore the effect of quality control circle (QCC) on health education in stroke rehabilitation nursing. Methods Such activities were conducted in the VIP ward of neurological rehabilitation in our hospital as: theme selecting, activity planning, status mastering, cause analyzing, countermeasures formulating and implementing, etc., in accordance with QCC theory from January to April, 2014. Results The satisfaction of stroke patients to health education rose from 80.1% to 91.1% (P<0.001), and the dissatisfaction to guidance of activities of daily living and prevention of complications decreased significantly (P<0.05) after the implementation of QCC program. All the circle members benefited from the improvement of QCC technique, team work, professional knowledge, communication and coordination, activity confidence, duty and honor. Conclusion Carrying out the QCC activities can play a positive role on health education in stroke rehabilitation nursing.

Objective To investigate the expression and the relationship with angiogenesis of miR-195 and NLR family member X1 (NLRX1) in granulation tissue after negative-pressure wound treatment (NPWT).Methods Six patients were collected who received negative pressure treatment with refractory wound granulation.The levels of miR-195, NLRX1 mRNA and NLRX1 proteins were measured.The expression of NLRX1 and the micro-vascular density (MVD) of CD31 were detected by immunohistochemistry (IHC).Results MiR-195 and MVD were significantly higher in granulation tissue after 7 days negative pressure treatment (P＜0.05), and NLRX1 was significantly lower (P ＜0.05).In granulation tissue,the expression of miR-195 was negatively correlated with NLRX1 (r =-0.856, P ＜0.001), the expression of NLRX1 was negatively correlated with MVD (r =-0.618, P ＜0.05), and the expression of miR-195 was positively correlated with MVD (r =0.630, P ＜ 0.05).Conclusions Negative pressure wound therapy can promote the formation of granulation vessels and the wound healing.The therapeutic mechanism may inhibit the expression of NLRX1 and upregulate the expression of miR-195 to promote angiogenesis.

Objective:To evaluate the clinical value of color Doppler ultrasound(CDUS) combined with vascular enhancement technology(VET) in diagnosis of iliac vein compression syndrome(IVCS).Methods:From Jan 2016 to Oct 2018, 252 patients with the lower extremities chronic venous diseases(CVD) were selected in the Second Hospital of Hebei Medical University. The ipsilateral iliac veins of the affected limbs were examined by CDUS, VET and the combined diagnosis of IVCS before X-Ray venography(XRV). Iliac vein diameter stenosis ratio(DSR)>50% in transverse section was the criterion of ultrasound diagnosis of IVCS. The stenosis site of iliac vein and indirect signs of IVCS, such as presence of collateral circulation and the retrograde flow of internal iliac vein were recorded. The cases, which had the same results in CDUS, VET and both and XRV, were divided into IVCS group and non-IVCS group. The results of XRV were taken as the gold standard, the diagnostic efficiency of the above 3 methods in diagnosis of IVCS was calculated. The cases identically diagnosed by the most effective ultrasonic method and XRV were divided into DVT group and non-DVT group according to the deep vein thrombosis in lower limbs. In the non-DVT group, there were five groups of C2-C6 on the basis of the CEAP clinical grades of CVD in lower extremity. The relationship between IVCS and different CEAP clinical grades were analyzed. The locations of common iliac vein stenosis and collateral circulation formation and internal iliac vein reverse flow were evaluated for the diagnosis of the IVCS.Results:①XRV diagnosis of IVCS was used as the gold standard. Compared with CDUS and VET alone, the sensitivity and specificity of CDUS combined with VET was the highest(all P<0.05). ②The distribution of DVT and non-DVT was significantly different in IVCS group and non-IVCS group diagnosed by the CDUS combined with VET(χ 2=145.0, P<0.001). ③In the non-DVT group, statistically significant differences of grades C3 and C5 were found between IVCS group and non-IVCS group(all P<0.05), while the differences of grades C2, C4 and C6 were non-significant(all P>0.05). ④For the proportion of the iliac vein stenosis sites, the prevalence of the primary section of left common iliac vein was much higher than those of the primary section of right common iliac vein and the middle-distal sections of bilateral common iliac veins(all P<0.05). There was no statistical difference between the middle-distal section of bilateral common iliac veins, and nor was bilateral junction area between external iliac vein and common femoral vein(all P>0.05). ⑤In IVCS group, which had the same results of CDUS combined with VET and XRV, there were statistical differences in the positive rate of collateral circulation and the retrograde flow of internal iliac vein(χ 2=6.717, P=0.010), and the former is higher than the latter. Conclusions:CDUS combined with VET has a higher diagnostic efficiency for IVCS than VET or CDUS alone. The presence of IVCS is closely related to DVT of lower extremities, but not related to clinical class of CEAP. The most common site of IVCS is the initial segment of the left common iliac vein. The presence of collateral circulation can be used as indirect indicators for the diagnosis of IVCS.