Objective To explore the effect of recombinant pEGFP-N3-APC vectors carrying various APC functional domains on the expression of β-catenin in human colorectal cancer cells HT-29.Methods The recombinant plasmids were transfected into HT-29 cells mediated by lipofectamine~(TM) 2000, and detected by green fluorescence and RT-PCR. Western blot was applied to detect β-catenin expression level in HT-29 cells after transfection, and gray scales of electrophoresis strips were analyzed by SPSS 13.0.Results Green fluorescence and RT-PCR made clear that all 5 recombinant plasmids were successfully expressed in HT-29 cells. Western blot showed that β-catenin expression level in HT-29 cells was not affected after being transfected with pEGFP-N3-APC1, pEGFP-N3-APC2 and pEGFP-N3-APC3, and was distinctly affected after being transfected with pEGFP-N3-APC4 and pEGFP-N3-APC5, especially the later one. Conclusion The selected APC5 gene fragment with 15-amino acid repeats and SAMP repeats, which is relatively short, can degrade β-catenin level in HT-29 cells and may be applied in the gene therapy.

Acupuncture Therapy ; Diaphragm ; pathology ; Humans ; Male ; Middle Aged ; Respiratory Paralysis ; pathology ; therapy

BACKGROUND:Animal and cel studies have shown that the Kidney Recipe can prevent and treat osteoporosis and improve bone metabolism, but this recipe is complicated. Recent studies on compound Chinese medicine mainly focused on serum drug metabolism and pharmacokinetics, which has limitations, and the effective ingredient and pharmaceutical material basis are uncertain. OBJECTIVE:In the different concentrations and time, by using different compatibility proportion of active ingredients of Kidney Recipe, osteoblasts from neonatal Sprague-Dawley rats were subjected to culture intervention. The proliferation and differentiation of osteoblasts were determined so as to identify time-effect and dose-effect relationship of Kidney Recipe on osteoblasts and to provide experimental evidences for prevention and treatment of osteoporosis. METHODS:Primary neonatal 24-hour osteoblasts of Sprague-Dawley rats were cultured in vitro. Herbs“tonic”and“cathartic”active chemical components of different proportion were used. The experiment contained three groups:Tonics Medicine (T)>Cathartic Medicine (C) group, TC group and TC group (P<0.05). At mass concentration of 20μg/L, no significant difference was detected in the T>C and TC group (P<0.05), and the proliferation became weak at 72 hours. At mass concentration of 10μg/L and 72 hours of culture, alkaline phosphatase secretion could be promoted in the T>C group. At mass concentration of 20μg/L and 24 hours of culture, the promoting effect on alkaline phosphatase secretion was most significant in the TC group (P<0.05). Results confirm that active ingredients of Kidney Recipe compatibility proportion can promote osteoblasts proliferation and differentiation most strongly in the T>C group in the time-effect and the dose-effect relationship.

As the third-generation bisphosphonate,zoledronic acid (ZOL) has been widely used for the treatment of skeletal-related events in breast cancer.Previous studies have demonstrated that ZOL has antitumor activitie in breast cancer.ZOL can induce direct and indirect antitumor activities through inducing apoptosis,inhibiting the proliferation,adhesion and invasion formation of bone microenvironment which is conducive to tumor growth,besides inhibiting angiogensis and modulating immune response.Also evidences of synergistic antitumor effects are found when ZOL combines with radiotherapy and chemotherapy,endocrine,and targeted drugs.The observed improvement in antitumor effects by using combination therapy with ZOL in breast cancer is currently being verified through additional clinical trials.

Objective To investigate the nursing points of the vacuum sealing drainage technology in children with refractory soft tissue infection in the limb.Methods A total of 9 children with refractory soft tissue infection in the limb were treated with vacuum sealing drainage technology from December 2010 to October 2012.Results All children who underwent vacuum sealing drainage technology were successful,among which 7 cases underwent continuous vacuum sealing drainage for 7 days and 2 cases replaced VSD material after continuous vacuum sealing drainage for 10 days and all wound with skin graft healed,without any complication.Conclusions Vacuum sealing drainage can remove thoroughly necrotic tissue of the wound and promote wound healing,the key points of nursing are to keep drainage tube unobstructed,adjust appropriate negative pressure and give close observation.

Retinal vein occlusion ( RVO ) is a common vascular disease of the retina and is one of the main reasons for blindness. ln recent years, there have been some new understanding about the diagnosis and treatment of the disease, especially some new researches about treatment,for example ,in the therapy of the intravitreal injection of triamcinolone acetonide and anti-VEGFs as well as dexamethasone implant ( Ozurdex ) . This article will make a brief summarization of the progress about the diagnosis and treatment of RVO.

ObjectiveTo observe the effects of electroacupuncture on IL-1β in brain tissue of rats after total cerebral ischemia-reperfusion.MethodsThirty male Wistar rats were randomly divided into the sham operation group, model group and acupuncture group, in each group. In acupuncture group and model group, 4-vessel-occlusion method was used to make a rat model of cerebral ischemia reperfusion. Acupuncture group used electroacupuncture on points Quchi and Zusanli, 6 hours after reperfusion. The rats of each group were killed and ELISA was used for the measurement of IL-1β in brain tissues.ResultsIL-1β in brain tissue in model group increased significantly compared with sham operation group and acupuncture group(P<0-001). No significant differences were found between sham operation group and acupuncture group (P>0-05).ConclusionAcupuncture can lower IL-1β content of brain tissue of rats with brain ischemia/reperfusion injury.

OBJECTIVETo explore the clinical application of mandibular-driven simultaneous maxillo-mandihular distraction to correct hemifacial microsomia with rapid prototyping technology.

METHODSThe patient' s skull resin model was manufactured with rapid prototyping technology. The osteotomy was designed on skull resin model. According to the preoperative design, the patients underwent Le Fort I osteotomy and mandibular ramus osteotomy. The internal mandible distractor was embedded onto the osteotomy position. The occlusal titanium pin was implanted. Distraction were carried out by mandibular-driven simultaneous maxillo-mandihular distraction 5 days after operation.

RESULTSThe distraction in five patients was complete as designed. No infection and dysosteogenesis happened. The longest distance of distraction was 28 mm, and the shortest distance was 16 mm. The facial asymmetry deformity was significantly improved at the end of distraction. The ocelusal plane of patients obviously improved.

CONCLUSIONSRapid prototyping technology is helpful to design precisely osteotomy before operation. Mandibular-driven simultaneous maxillo-mandibular distraction can correct hemifacial microsomia. It is worth to clinical application.

Face ; abnormalities ; surgery ; Facial Asymmetry ; congenital ; surgery ; Goldenhar Syndrome ; surgery ; Humans ; Hyperplasia ; surgery ; Mandible ; surgery ; Maxilla ; surgery ; Osteogenesis, Distraction ; methods ; Osteotomy ; methods ; Osteotomy, Le Fort

Objective To construct recombinant plasmids containing various functional domains of APC protein and detect their expression in HCT-116 cells. Methods Five APC gene fragments were amplified by PCR with whole APC gene as template and primers designed according to APC cDNA sequence and mutation cluster domain. The five obtained fragments were cloned into eukaryotic expression vector pEGFP-N3 to generate recombinant pEGFP-N3-APC1-5. Sequence of the inserted gene was identified and analyzed after restriction enzyme digestion. Liposome-mediated recombinant plasmid pEGFP-N3-APC was transfected into HCT 116 cells and identified by green fluorescence. RT-PCR was employed to validate the expression of recombinant vectors in cells. Results Recombinant pEGFP-N3-APC1-5 were confirmed by restriction enzyme digestion and sequence analysis. The plasmids could be expressed in HCT-116 cell line detected by fluorescence microscope. Results of RT-PCR made clear that vectors constructed could be expressed in HCT-116 cells. Conclusion The relative efficient expression of five recombinant expressive vector in HCT-116 cell line may provide an experimental basis for selecting specific therapy peptide for colorectal cancer.

In this study, UPLC-MS/MS was adopted to determine the contents of five ephedrine alkaloids (Norephedrine, Norpseudoephedrine, Ephedrine, Pseudoephedrine, Methylephedrine) in plasma and urine in rats after the combined administration of Ephedrae Herba-Gypsum Fibrosum and calculate relevant pharmacokinetic parameters, in order to discuss the effect of the combined administration of Ephedrae Herba-Gypsum Fibrosum on plasma pharmacokinetics and urinary excretion characteristics. According to the results, after being combined with Gypsum, the five ephedrine alkaloids showed similar pharmacokinetic changes, such as shortened t(max), accelerated absorption rate, but reduced AUC(0-t) and V(z)/F, which may be related to the increase in urine excretion. Besides, Gypsum was added to enhance C(max) of Pseudoephedrine and prolong MRT(0-t) of Methylephedrine, so as to enhance the anti-asthmatic effect of Ephedrae Herba and resist the toxic effect of Norephedrine and Ephedrine. This study proved the scientific compatibility of Ephedrae Herba-Gypsum Fibrosum and provided a reference for studies on the prescription compatibility regularity and relevant practices.
Alkaloids ; blood ; pharmacokinetics ; urine ; Animals ; Calcium Sulfate ; pharmacokinetics ; Drugs, Chinese Herbal ; pharmacokinetics ; Ephedra ; chemistry ; Male ; Rats ; Rats, Sprague-Dawley ; Urine ; chemistry