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1.
Frontiers of Medicine ; (4): 105-118, 2023.
Article in English | WPRIM | ID: wpr-971622

ABSTRACT

The third-generation epidermal growth factor receptor (EGFR) inhibitor osimertinib (OSI) has been approved as the first-line treatment for EGFR-mutant non-small cell lung cancer (NSCLC). This study aims to explore a rational combination strategy for enhancing the OSI efficacy. In this study, OSI induced higher CD47 expression, an important anti-phagocytic immune checkpoint, via the NF-κB pathway in EGFR-mutant NSCLC HCC827 and NCI-H1975 cells. The combination treatment of OSI and the anti-CD47 antibody exhibited dramatically increasing phagocytosis in HCC827 and NCI-H1975 cells, which highly relied on the antibody-dependent cellular phagocytosis effect. Consistently, the enhanced phagocytosis index from combination treatment was reversed in CD47 knockout HCC827 cells. Meanwhile, combining the anti-CD47 antibody significantly augmented the anticancer effect of OSI in HCC827 xenograft mice model. Notably, OSI induced the surface exposure of "eat me" signal calreticulin and reduced the expression of immune-inhibitory receptor PD-L1 in cancer cells, which might contribute to the increased phagocytosis on cancer cells pretreated with OSI. In summary, these findings suggest the multidimensional regulation by OSI and encourage the further exploration of combining anti-CD47 antibody with OSI as a new strategy to enhance the anticancer efficacy in EGFR-mutant NSCLC with CD47 activation induced by OSI.


Subject(s)
Humans , Mice , Animals , Carcinoma, Non-Small-Cell Lung/metabolism , Lung Neoplasms/metabolism , Acrylamides/pharmacology , ErbB Receptors/metabolism , Cell Line, Tumor , CD47 Antigen/therapeutic use
2.
Acta Pharmaceutica Sinica B ; (6): 1467-1487, 2023.
Article in English | WPRIM | ID: wpr-982795

ABSTRACT

Described as a "don't eat me" signal, CD47 becomes a vital immune checkpoint in cancer. Its interaction with signal regulatory protein alpha (SIRPα) prevents macrophage phagocytosis. In recent years, a growing body of evidences have unveiled that CD47-based combination therapy exhibits a superior anti-cancer effect. Latest clinical trials about CD47 have adopted the regimen of collaborating with other therapies or developing CD47-directed bispecific antibodies, indicating the combination strategy as a general trend of the future. In this review, clinical and preclinical cases about the current combination strategies targeting CD47 are collected, their underlying mechanisms of action are discussed, and ideas from future perspectives are shared.

3.
International Eye Science ; (12): 1079-1084, 2022.
Article in Chinese | WPRIM | ID: wpr-929483

ABSTRACT

AIM: To investigate the effect of modified Zhujing pill on retinal autophagy in mice with form deprivation myopia.METHODS: Thirty C57BL/6 mice were randomly divided into a negative control group, a myopia model group and a traditional Chinese medicine intervention group, with 10 mice in each group. Except for the negative control group, all mice in the myopia model group and the traditional Chinese medicine intervention group used translucent EP tubes to cover their right eyes to make a form deprivation myopia(FDM)model; The traditional Chinese medicine intervention group gavage Zhujing pill modified suspension 0.546g/(kg·d)(0.15mL/d), the negative control group and the myopia model group were given an equal amount of normal saline(0.15mL/d)for 4wk. At the beginning and the end of the experiment respectively, the right eye diopter of the mouse was measured with a strip retinoscope, measurement of the axial length of the right eye of mouse by A-ultrasound. At the end of the experiment, the right eyes of all mice were taken for detection, and immunofluorescence method was used to locate and detect the activity and migration of the retinal microglia marker(Iba1); Transmission electron microscope observation of autophagosome formation in retinal pigment epithelial cells; Western Blot, real-time fluorescent quantitative PCR(q-PCR)to detect the autophagy marker LC3Ⅱ and p62 protein quantitative and gene expression in retinal tissues.RESULTS: At the end of the experiment, the refractive power of the right eyes of mice showed that the myopia model group and the traditional Chinese medicine intervention group formed relative myopia, the myopia model group and the traditional Chinese medicine intervention group were significantly lower than those of the negative control group(all P<0.01). At the end of the experiment, the axial length of the myopia model group and the Chinese medicine intervention group were significantly increased compared with the negative control group(P<0.01). Immunofluorescence method for locating and detecting Iba1 showed that the average optical density of Iba1 in the retina of the myopia model group increased the most obviously, followed by the increase in the negative control group, and the decrease in the traditional Chinese medicine intervention group. Compared with the negative control group, the myopia model group increased significantly(P<0.05), and the traditional Chinese medicine intervention group was significantly lower than the myopia model group(P<0.05). It was found that Iba1 migrated to the ganglion cell layer in the myopia model group and the traditional Chinese medicine intervention group. Transmission electron microscopy showed that autophagosomes were observed in the retinal pigment epithelial cells of the myopia model group and the Chinese medicine intervention group. The results of Western Blot and q-PCR showed that the expression of LC3Ⅱ and p62 increased most obviously in the traditional Chinese medicine intervention group, followed by the myopia model group, and the negative control group was the lowest.CONCLUSION: The results of the study show that modified Zhujing pill may enhance retinal autophagy in mice with FDM by inhibiting the activation of microglia.

4.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 196-203, 2022.
Article in Chinese | WPRIM | ID: wpr-940569

ABSTRACT

ObjectiveTo investigate the potential pharmacological mechanism of Xinmaikang tablets in the treatment of atherosclerosis cardiovascular disease by using network pharmacology and cell experimental validation. MethodThe components of Xinmaikang tablets were searched by BATMAN-TCM database and the active ingredients and potential targets were screened. The atherosclerosis related disease targets were searched in GeneCards and online mendelian inheritance in man(OMIM) disease databases. The therapeutic targets were obtained by mapping the intersection of the tablets and disease targets. Therapeutic targets were uploaded to STRING database to construct protein-protein interaction(PPI) network. Cytoscape software was used to create a "drug-active component-therapeutic target" network map, and a network topology algorithm was used to screen key action targets. David software was used for gene ontology(GO) and Kyoto encyclopedia of genes and genomes(KEGG) function enrichment analysis. The key targets of drug therapy were validated by in vitro cell assay. ResultA total of 19 active ingredients, 132 potential targets and 4703 atherosclerotic disease-related target genes of Xinmaikang tablets were retrieved and screened, and 84 intersection targets were obtained. 3 key therapeutic targets of Xinmaikang tablets in the treatment of atherosclerotic diseases were screened, including Calmodulin 1(CALM1), voltage-dependent L-type calcium channel subunit alpha-1C(CACNA1C) and Phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha isoform(PIK3CA). A total of 313 biological processes, 89 molecular functions and 53 cell components were obtained by GO enrichment. A total of 40 pathways were obtained from KEGG functional enrichment, including purine metabolism, renin secretion, CGMP/PKG signaling pathway and so on. In vitro cell experiment results verified that Xinmaikang tablets can up-regulate the expression of CALM1 and CACNA1C, down-regulate the expression of PIK3CA, so as to inhibit the activity of inflammatory response, and play a therapeutic role in atherosclerotic diseases. ConclusionXinmaikang tablets may treat atherosclerosis cardiovascular disease through betulin, methyleugenol and other compounds, through purine metabolism, renin secretion, cGMP/PKG signaling pathway and other pathways, which acts on CALM1, CACNA1C, PIK3CA and other targets.

5.
China Journal of Orthopaedics and Traumatology ; (12): 554-558, 2021.
Article in Chinese | WPRIM | ID: wpr-888314

ABSTRACT

OBJECTIVE@#To investigate the clinical effect of double mini-locking plates in the treatment of ulna olecranon fractures.@*METHODS@#From March 2017 to May 2020, 19 patients with olecranon fractures were treated with double mini locking plates, including 12 males and 7 females, aged from 20 to 75 years old with an average of (40.50±7.62) years old;10 patients had the injuries on the left side and 9 patients on the right side. All the 19 patients were fresh closed fractures without ulnar coronoid process fracture, elbow dislocation and other injuries. The fracture healing time and complications were recorded, and the clinical efficacy was evaluated by Mayo elbow performance score (MEPS) before operation and 12 months after operation.@*RESULTS@#All the 19 patientswas followed up, and the duration ranged from 12 to 17 months with an average of (13.51±3.17) months. Postoperative follow-up showed all fractures healed. Fracture healing time ranged from 2 to 6 months, with an average of(3.77±1.24) months. There was no internal fixation fracture, screw loosening, infection, internal fixation irritation, heterotopic ossification, elbow stiffness and other complications occurred. The MEPS score of affected elbow at 12 months after operation was 91.26±3.87, which was significantly different from that before operation 56.18±9.56 (@*CONCLUSION@#It is a reliable fixation method to treat olecranon fracture with double mini locking plate. The incision lengh is small and the fracture fixation is reliable. Elbow joint function exercise can be performed early after operation. Postoperative internal fixation has less skin irritation and satisfactory elbow joint function recovery.


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Bone Plates , Elbow Joint , Fracture Fixation, Internal , Olecranon Process , Treatment Outcome , Ulna Fractures
6.
Chinese Journal of Tissue Engineering Research ; (53): 1452-1460, 2016.
Article in Chinese | WPRIM | ID: wpr-485672

ABSTRACT

BACKGROUND:Stem cel therapy has been used for prevention and treatment of degenerative disc disease. Considering the special microenvironment in the intervertebral disc, the survival rate and differentiation ability of transplanted cels are decreased, which may lead to the poor efficacy of stem cel therapy. How to improve the survival ability and therapeutic effect of the transplanted cels is the focus of stem cel therapy for degenerative disc disease. OBJECTIVE: To investigate the effects of cobalt chloride combined with hypertonic solution pretreatment on bone marrow mesenchymal stem cels that wil be transplanted for treatment of degenerative disc disease. METHODS:(1)In vitro cel experiment: bone marrow mesenchymal stem cels were divided into three groups and subjected to normal culture medium (normal control group), 1% hypertonic mother solution (hypertonic group), 100 μmol/L cobalt chloride (hypoxia group), or 1% hypertonic mother solution plus 100 μmol/L cobalt chloride (combined group) for 1 week. Then, 2% hypertonic solution and 200 μmol/L cobalt chloride cobalt chloride were used to simulate the anaerobic and hypertonic environment intervenes in pretreated and untreated bone marrow mesenchymal stem cels for 24 hours. After that, RT-PCR was used to detect the expression of caspase-3 for apoptosis evaluation. (2)In vivo animal experiment: Sprague-Dawley rats were divided into model, cel transplantation and hypertonic plus hypoxic groups. Rat models of intervertebral disc degeneration were made in these three groups. After modeling, rats in these three groups were given no treatment, bone marrow mesenchymal stem cel transplantation or transplantation of bone marrow mesenchymal stem cels which were subjected to hypertonic and hypoxia pretreatments into the intervertebral disc. Two weeks later, immunohistochemistry and RT-PCR methods were used to detect cel distribution and related gene expression, respectively, thereby to evaluate the therapeutic effect of stem cels. RESULTS AND CONCLUSION: (1)In vitro cel experiment: caspase-3 mRNA expression was significantly reduced in pretreated bone marrow mesenchymal stem cels compared with the untreated cels (P < 0.05). (2)In vivo animal experiment: compared with the control group, the caspase-3 and interleukin-1β in the intervertebral disc and a number of degenerative indexes were decreased in the cel transplantation. Compared with the cel transplantation group, these indicators had better outcomes in the hypertonic plus hypoxic group (P < 0.05). These findings indicate that bone marrow mesenchymal stem cels have therapeutic potential for degenerative disc disease, and have better adaptability and transplantation effects by hypertonic and hypoxia pretreatments.

7.
Chinese journal of integrative medicine ; (12): 733-742, 2015.
Article in English | WPRIM | ID: wpr-287156

ABSTRACT

<p><b>OBJECTIVES</b>To investigate the safety and efficacy of yangxinkang tablets in patients with chronic heart failure (CHF) and syndrome of qi and yin deficiency, blood stasis, and water retention.</p><p><b>METHODS</b>In a double-blinded, randomized, placebo-controlled, multicenter clinical trail, 228 patients with CHF New York Heart Association (NYHA) class II or III in stage C were assigned by randomized block method to two groups in a 1:1 ratio to undergo either conventional Western treatment or conventional treatment plus yangxinkang tablets for 4 weeks. The outcome measure were effect of cardiac function, Chinese medicine (CM) syndromes, scores of symptoms, signs, and quality of life measured by Minnesota Living with heart failure questionnaire (MLHFQ) before and after the treatment.</p><p><b>RESULTS</b>Totally 112 patients were analyzed in the treatment group and 109 in the control group. They were comparable in NYHA functional class, basic parameters and primary diseases before treatment. Cardiac function and CM syndromes were greatly ameliorated in both groups after treatment. Total effective rates of cardiac function and CM syndrome in the treatment group were significantly higher than those in the control group (P<0.05). Total symptom score and sign score in the treatment group decreased significantly after treatment (P<0.01), which were significantly lower than those in the control group (P<0.05). There were statistically significant differences in post-treatment scores of gasp, cough with phlegm, pulmonary rales and jugular vein engorgement between the two groups (P<0.05 or P<0.01). Three MLHFQ scores decreased significantly in both groups after treatment (P<0.01). Post-treatment total scale score and physical subscale score in the treatment group and the reduction of them showed statistically significant differences (P<0.05) as compared with the control group. There was no significant difference between the two groups in emotional subscale score and the reduction after treatment (P>0.05). There was no obvious adverse reaction in either group noted during the study.</p><p><b>CONCLUSIONS</b>Yangxinkang tablets were safe and efficacious in improving cardiac function, CM syndromes, symptoms, signs, and quality of life in patients with CHF class II or III in stage C on the base of conventional treatment.</p>


Subject(s)
Aged , Female , Humans , Male , Chronic Disease , Double-Blind Method , Drugs, Chinese Herbal , Heart Failure , Drug Therapy , Quality of Life , Surveys and Questionnaires , Tablets
8.
Chinese Journal of Virology ; (6): 238-245, 2014.
Article in Chinese | WPRIM | ID: wpr-356609

ABSTRACT

This study aims to investigate the virological impact of the stalk region and cysteine (C) in neuraminidase (NA) of influenza A/Anhui/1/05 (H5N1) and A/Ohio/07/2009 (H1N1) viruses. The NA of A/ Anhui/1/05 (H5N1), defined as AH N1, lacked 20 amino acids (including C, defined as s20) as compared with NA of A/Ohio/07/2009 (H1N1) (defined as 09N1). We deleted s20 of 09N1 to construct 09N1-s20, and inserted s20 into AH N1 to construct AH N1+s20. To investigate the impact of C on the biological function of NA, we deleted C in 09N1 to construct 09N1-C and inserted C into AH N1 to construct AH N1-C. The pseudo-type viral particle (pp) system was used to evaluate the impact of these mutants on virology. The combination of 09N1-C and 09H1 (defined as 09H1::09N1-C) showed an infectivity 8 times that of the wild type 09H1::09N1, while the infectivity of the combination of AH N1+C and AH H5 (defined as AH H5::AH N1+C) was much lower than that of the wild type AH H5::AH N1. The infectivity of the combination of 09N1-s20 and 09H1 (defined as 09H1::09N1-s20) was 4 times that of the wild type 09H1::09N1; the infectivity of the combination of AH N1+s20 and AH H5 (defined as AH H5:: AH N1+s20) was 1/7 that of the wild type AH H5::AH N1. The co-existence of 09N1-C and AH H5 displayed 6 times the infectivity of AH H5::09N1, while the infectivity of 09H1::AH N1+C was very low. Multimer analysis showed that in the wild type 09N1, the forms of NA were dimer > tetramer > monomer; the major component of NA in 09N1-C was monomer; in 09N1-s20, the forms of NA were monomer > dimer. AH N1 was mainly composed of monomer; in AH N1+s20, the forms of NA were dimer > monomer > tetramer; in AH N1+C, the forms of NA were dimer > tetramer. Deletion of C or s20 from 09N1 did not change the expression of NA. The study suggested that deletion of C from the stalk region of NA in A/Ohio/07/2009 (H1N1) increases infectivity. Insertion of C into NA's stalk region of A/ Anhui/1/05 (H5N1) significantly decreases infectivity. Cysteine deletion in the stalk region is important for the infectivity of A/Anhui/1/05 (H5N1) and A/Ohio/07/2009 (H1N1). It may interfere with the infectivity via changes in NA polymerization.


Subject(s)
Humans , Amino Acid Motifs , Influenza A Virus, H1N1 Subtype , Chemistry , Genetics , Virulence , Influenza A Virus, H5N1 Subtype , Chemistry , Genetics , Virulence , Influenza, Human , Virology , Neuraminidase , Chemistry , Genetics , Metabolism , Viral Proteins , Chemistry , Genetics , Metabolism , Virulence
9.
China Journal of Orthopaedics and Traumatology ; (12): 651-653, 2012.
Article in Chinese | WPRIM | ID: wpr-321892

ABSTRACT

<p><b>OBJECTIVE</b>To investigate therapeutic effects of vacuum sealing drainage (VSD) in the treatment of soft tissue defect combined with tendon and bone exposure.</p><p><b>METHODS</b>From October 2007 to February 2011, 397 patients (412 feet) with open ankle fracture and dislocation combined with soft tissue defected were treated by VSD. There were 301 males and 96 females with an average age of 36 years (ranging age from 20 to 73 years). According to AO classification, 74 feet were type I, 211 feet were type II, 108 feet were type III and 19 feet were type IV. The mean time from injury to operation was 5.6 h ( 2 to 12 h). The mean treatment time of was 10 months (4 to 19 months).</p><p><b>RESULTS</b>One hundred and forty-one patients were primarily healed, 97 patients were sutured at stage II. Split-thick skin grafting was performed at stage II was performed in 103 patients; free flap transplantation was performed in 25 patients. Three of the 34 patients with infection were removed steel plate; Eviscerate flap coverage wound was performed in 14 patients caused by the first metatarsal bone exposure; Toe amputation were performed in 22 cases caused by toes necrosis. Tarsometatarasl joints perforators' surgery was performed in 10 patients with forefeet necrosis. Thirty hundred and six patients were followed up from 3 to 20 months (averaged 10 months). The wounds healed well.</p><p><b>CONCLUSION</b>VSD for soft tissue defects caused by ankle injury is a simple and effective method, but can not replace debridement and transfer flap.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Ankle Fractures , Debridement , Drainage , Methods , Joint Dislocations , General Surgery , Skin Transplantation , Treatment Outcome , Vacuum
10.
China Journal of Orthopaedics and Traumatology ; (12): 645-647, 2011.
Article in Chinese | WPRIM | ID: wpr-351650

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the clinical results of the proximal femoral nail antirotation (PFNA) system in the treatment of unstable intertrochanteric femoral fractures.</p><p><b>METHODS</b>From September 2006 to September 2009, 90 patients (40 males and 50 females, ranged in age from 64 to 95 years with an average of 73.2 years with unstable intertrochanteric femoral fractures were surgically treated with PFNA. Fifty patients had the fractures in the right hip, and 40 patients had the fractures in the left hip. The fractures were classified according to the AO classification: 11 patients were type A2.1, 21 patients were type A2.2,25 patients were type A2.3 9 patients were type A3.1,6 patients were A3.2 and 18 patients were A3.3. The patients underwent surgery within a mean of 3.2 days(ranged,2 to 20.1 days) from injury. The mean hospital stay was 12.8 days(ranged,7 to 24 days). Closed reduction was achieved in all the patients. Harris hip score were used for the evaluation of clinical effects.</p><p><b>RESULTS</b>The mean operation time was 36.8 min (ranged, 23 to 110 min) and the mean blood loss was 150 mi (ranged, 100 to 500 ml). The mean follow-up period was 12 months (ranged, 6 to 24 months). All the patients had fracture union. Sixty-nine patients got excellent reduction, 14 good and 7 bad. The mean collodiaphysial angle was 135.60 (ranged, 1260 to 1470). Postoperative complications included secondary varus in 2 patients,calcification at the tip of the greater trochanter in 5 patients, medial thigh pain in 7 patients,and screw cut-out in 1 petient. Ten patients had femoral shortness (mean 9.3 mm,ranging from 8 to 14 mm). The mean Harris hip score was (80.5 +/- 9.8). According to Harris hip scores evaluation system, 26 patients reached an excellent result, 37 good, 18 poor and 9 bad.</p><p><b>CONCLUSION</b>Due to advantages of high union rate, short operation time, and early postoperative mobilization, PFNA osteosynthesis is an idea method for surgical treatment of unstable intertrochanteric femoral fractures.</p>


Subject(s)
Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Bone Nails , Fracture Fixation, Internal , Methods , Hip Fractures , Diagnostic Imaging , General Surgery , Radiography
11.
Journal of Southern Medical University ; (12): 979-982, 2011.
Article in Chinese | WPRIM | ID: wpr-332503

ABSTRACT

<p><b>OBJECTIVE</b>To explore the effects of serum of asthmatic patients, dexamethasone, interleukin-4 (IL-4), interferon-gamma (IFN-γ) and transforming growth factor-β (TGF-β) on the expression of interleukin-22 receptor 1 (IL-22R1) mRNA and protein in HASMCs in vitro.</p><p><b>METHODS</b>IL-22R1 mRNA and protein expressions in HASMCs treated with different stimulating agents were measured by real-time PCR and Western blotting, respectively.</p><p><b>RESULTS</b>IL-22R1 mRNA and protein expressions in HASMCs were significantly increased after stimulation by serum from asthmatic patients, but decreased after co-stimulation with dexamethasone. IL-22R1 mRNA and protein expressions in the cells both increased after stimulation by IL-4, IFN-γ and TGF-β.</p><p><b>CONCLUSION</b>IL-22R1 in HASMCs might be involved in the pathogenesis of asthma, and the therapeutic effect of dexamethasone on asthma is mediated, at least partially, by IL-22R1. The effects of IFN-γ, IL-4, and TGF-β on asthma may also be attributed to their actions on HASMCs.</p>


Subject(s)
Humans , Asthma , Blood , Cell Line , Interferon-gamma , Pharmacology , Interleukin-4 , Pharmacology , Myocytes, Smooth Muscle , Metabolism , RNA, Messenger , Genetics , Receptors, Interleukin , Metabolism , Transforming Growth Factor beta , Pharmacology
12.
Journal of Southern Medical University ; (12): 1102-1105, 2009.
Article in Chinese | WPRIM | ID: wpr-282610

ABSTRACT

<p><b>OBJECTIVE</b>To observe the effect of exogenous phophatase and tensin homolog deleted on chromosone 10 (PTEN) gene transfer via recombinant adenoviruses on the proliferation of human airway smooth muscle cells (HASMCs) in vitro and investigate the possible mechanisms.</p><p><b>METHODS</b>With a recombinant adenovirus vector containing PTEN (Ad-PTEN) constructed using the pAdxsi system, PTEN gene was transiently transfected into HASMCs and the transfection efficiency was determined by fluorescence microscope. RT-PCR and Western blotting were performed to detect the expression of PTEN mRNA and protein in the infected cells. MTS/PMS assay was used to analyze the proliferation of HASMCs, and the cell cycle changes of the transfected cells were evaluated by flow cytometry with PI staining. The expression levels of Akt and p-A kt proteins were detected by Western blotting, and P21 mRNA expression determined by RT-PCR.</p><p><b>RESULTS</b>The recombinant adenovirus Ad-PTEN showed a wild-type PTEN gene transfer efficiency of 98% at the multiplicity of infection (MOI) of 100. RT-PCR and Western blotting showed that infection with the recombinant adenovirus resulted in PTEN overexpression in the HASMCs, causing also increased ratio of G(0)/G(1) cells and proliferation inhibition of the ASMCs. The overexpression of PTEN significantly decreased the expression level of p-Akt but increased P21 mRNA expression.</p><p><b>CONCLUSION</b>The recombinant adenovirus containing PTEN can be successfully transfected into HASMCs cultured in vitro, resulting in PTEN overexpression at both the mRNA and protein levels. PTEN overexpression can efficiently inhibit the proliferation of HASMCs possibly through the PI3K/PKB/AKt and P21 pathways.</p>


Subject(s)
Humans , Adenoviridae , Genetics , Metabolism , Bronchi , Cell Biology , Cell Proliferation , Cells, Cultured , Genetic Vectors , Genetics , Muscle, Smooth , Cell Biology , PTEN Phosphohydrolase , Genetics , RNA, Messenger , Genetics , Recombinant Proteins , Genetics , Pharmacology , Transfection
13.
Journal of Southern Medical University ; (12): 859-863, 2009.
Article in Chinese | WPRIM | ID: wpr-268826

ABSTRACT

<p><b>OBJECTIVE</b>To construct a recombinant adenovirus vector containing phosphatase and tensin homolog deleted on chromosome 10 (PTEN) using the pAdxsi system.</p><p><b>METHODS</b>PTEN cDNA from plasmid pcDNA3-PTEN was cloned into the shuttle plasmid pShuttle-GFP-CMV. The shuttle vector was transformed into competent DH5alpha strain with the vector pAdxsi to achieve the homologous recombination. The recombinant construct was subsequently linearized with PacI and transfected into HEK293 cells via Lipofectamine 2000. The recombinant adenovirus particles were collected, and after titration, the recombinant adenovirus was traced by monitoring GFP expression under fluorescence microscope. The expression of PTEN mRNA and protein in the recombinant adenovirus vector and airway smooth muscle cells were detected by PCR and Western blotting, respectively.</p><p><b>RESULTS</b>GFP was expressed in HEK293 cells infected by recombinant adenovirus, and the expression intensity increased gradually with the passage of time, with obvious cytopathic effect (CPE) noted in the cells. After 3 cycles of amplification, the titer of adenovirus containing PTEN reached an appropriate level. The viral titer of pAdxsi-GFP-PTEN was 2x10(10) pfu/ml, and PTEN mRNA expression was detected by PCR. The homologous protein expressed in the infected human airway smooth muscle cells significantly increased in comparison with that in the control cells.</p><p><b>CONCLUSION</b>The recombinant adenovirus containing PTEN is constructed successfully, which provides an experimental basis for studying the role of PTEN gene in asthma therapy.</p>


Subject(s)
Humans , Adenoviridae , Genetics , Metabolism , Bronchi , Cell Biology , Cells, Cultured , Genetic Vectors , Genetics , Green Fluorescent Proteins , Genetics , Muscle, Smooth , Cell Biology , Metabolism , PTEN Phosphohydrolase , Genetics , RNA, Messenger , Genetics , Recombinant Fusion Proteins , Genetics , Transfection
14.
China Journal of Chinese Materia Medica ; (24): 738-740, 2005.
Article in Chinese | WPRIM | ID: wpr-358121

ABSTRACT

<p><b>OBJECTIVE</b>To establish an FTIR method for the analysis of Dendrobium.</p><p><b>METHOD</b>Using fourier transform infrared spectrometer to record the characteristic spectra of eleven samples of Dendrobium, and to compare the spectra by PCA (principal component analysis).</p><p><b>RESULT</b>The FTIR spectra of the upper part of the stem displayed significant differences between fresh and dried samples of Dendrobium. On the other hand, differences were observed in the spectra of the middle and lower parts of stems of D. guangxieuse when compared to other species.</p><p><b>CONCLUSION</b>The method of applying PCA to FTIR analysis is a rapid and dependable method for comparing samples of Dendrobium.</p>


Subject(s)
Dendrobium , Chemistry , Classification , Plant Stems , Chemistry , Plants, Medicinal , Chemistry , Classification , Principal Component Analysis , Methods , Spectroscopy, Fourier Transform Infrared , Methods
15.
Chinese Journal of Rehabilitation Theory and Practice ; (12): 434-434, 2004.
Article in Chinese | WPRIM | ID: wpr-979106

ABSTRACT

@#Objective To investigate the value of arthrolysis in treatment of scapulohumeral periarthritis.Methods 83 cases of scapulohumeral periarthritis were divided into arthrolysis group(45 cases), which were treated with arthrolysis combined with electrotherapy, and control group (38 cases),which were treated with electrotherapy only.Results The effect of arthrolysis group was better than that of control group (P<0.05).Conclusion Arthrolysis can improve the effect on scapulohumeral periarthritis.

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