Objective To investigate the effect of adenovirus-medicated gene therpy with vascular endothelial growth factor(VEGF)delivered into the subdermal space to treat compromised skin flap in the rat.Methods Thir- ty sprague-dawley rats weighing 350～400g were used in this study.The rats was randomly divided into three groups of ten rats each.A dorsal flap(8cm?2cm)in full thickness with the pedicale located at the leve of the iliaccrest was designed,each,animal received recipient bed subfascial injections of adenovirus encoding VEGF(Ad-VEGF)or green fluorescent protein(Ad-GFP)as treatment control.Another set of animals(n = 10)received NG was designated as control.The flap were elevated as originally designated and sutured back to its bed.The survival rate of the flap was evaluated on day seven after operation.Results The survival rates of the flap in Ad-VEGF group increased signifi- cantly as compared with those of the Ad-GFP group(P

AIM:To be one of the primary cause injury to multiple sites of ocular of glaucoma which affects over 70 million people worldwide. We applied data mining techniques, linear and the matrix operations, efficiently calculated the network and estimated the possible function of the“node” genes of the retina and optic of glaucoma, in order to provide new thought and method on the pathogenesis of glaucoma.
METHODS: The data in this study is from Gene Expression Omnibus ( GEO ) which belong to Nation Center for Biotechnology Information ( NCBI) , the quality of the raw data CEL files was processed and analyzed by the Expression software which belong to Affymetrix Inc. , Santa Clare, CA, USA. Significant analysis method ( SAM) which base on the T test was used to identified the significant genes. Based on GRNInfer and Gvedit soft we set up gene networks of optic and retina of mice and further more enriched analysis which based on DAVID and MAS3. 0 online software were processed.
RESULTS:The analysis between the group of the optic nerve heads and retinas in different stage of glaucoma showed that the amount of significant different expressed genes in the optic never head group increased significantly comparing with the group of retina in the early stage of glaucoma, the analysis of the genes network construction show that:the node genes of optic nerve heads included Unc13c、Kif5a、TRPM1、PANX; and the node genes of retina include POU4F1, NEFL, BC03870, CALB2. Metabolic pathways enrichment analysis which based on MAS3. 0 online platform show that there was mainly the amyotrophic lateral sclerosis, tyrosine metabolism, melanogenesis, Nitrogen metabolism, Gap junction, Leukocyte transendothelial migration metabolism pathway enriched out in optic nerve head; and there was mainly amyotrophic lateral sclerosis, neurodegenerative disorders, prostate cancer, leukocyte transendothelial migration metabolism pathway enriched out in retina.
CONCLUSION:By understanding bioinformatics result, it seems optic were more sensitive than the retina to high intraocular pressure, and weather high expression of TYrp1 gene can be as a sensitive diagnostic item require more evidence back up. Functional enrich analysis of node gene showed that cytoskeleton reconstructed, molecular motor and nutrients transport function improve in optic; and in retina, the most prominent finding in retina was enrichment function modules were focus on regeneration, repairing and differentiation of cells, which remind that we should reinforce research on reparation of retina of primary glaucoma. Metabolic pathways enrichment analysis show that inflammatory response plays prominent place in optic and retina of primary glaucoma, because of the optic narrow and crowed anatomic shape, nutrient metabolism and substances transfer enrichment modules play an important role in optics of primary glaucoma.

OBJECTIVETo see the change of capillary of heart in Athlete's Heart, so that to discover the mechanism of pathologic change.

METHOD18 male SD rats were separated randomly into control group (without any exercise), aerobic exercise group (swimming for 75 min every day), and overload group (swimming for 180 min with 5% weight of its body every day). After 5 days per week, 12 weeks, exercise training stopped and heart of rats were observed under Transmission Electron Microscope.

RESULTSIn aerobic exercise group, the capillary cavities in heart expand, the walls of capillary become thick; the number of mitochondrion increases; endothelium cells become active in growth. However, after overload exercise, the walls of capillary cockle and protuberances appear. The mitochondrion swell and the cristae become disorder. Most of endosomes expand and their number increases. The karyons become abnormity in shape and uniformity in electronic density, besides the nuclear envelope cockle. The basilar membranes become thick and unclear.

CONCLUSIONAfter exercise training, both physical and pathologic changes in heart capillary are found. In suitable exercises group, the capillaries change physically; the pathologic changes are becoming visible after overload exercise however.

Animals ; Capillaries ; ultrastructure ; Cardiomegaly ; etiology ; pathology ; Male ; Physical Conditioning, Animal ; adverse effects ; Physical Endurance ; Rats ; Rats, Sprague-Dawley ; Sports

ObjectiveTo explore the prevalence and spectrum of β-thalassemia mutations in Fujian province,and to provide a reference for prenatal diagnosis and genetic counseling in this population.Methods Two thousand three hundred and one blood samples were randomly selected from 9 different areas of Fujian province from May 2008 to December 2010.PCR and reverse dot blot hybridization (RDB) were adopted for detection of the 17 common types of mutation,and the frequency of each genotype of β-thalassemia mutations was calculated.The β-globin gene of unknown positive samples were analyzed directly with DNA sequencing.Results Three hundred and fifty-nine cases were detected with β-thalassemia mutations of the 2301 copy blood samples submitted,and the detection rate was 15.60％ (359/2301).Of the mutated genes,12 different mutations were identified,namely IVS-2-654(C→T),CD41-42(-TCTT),CD17(A→T),-28(A→G),CD27-28(+C),CD26(G→A),CD71-72(+A),IVS-1-1(G→T),CD43(G→T),-29(A→G),initiation codon ATG→AGG and CD36(-C).Mutation frequencies were 46.54％ (175/376),33.24％ (125/376),9.31％ (35/376),5.05％ (19/376),2.13％(8/376),1.33％(5/376),0.80％(3/376),0.27％(1/376),0.27％(1/376),0.27％(1/376),0.53％(2/376),and 0.27％(1/376),respectively.The most common mutations were IVS-2-654 (C→T) and CD41-42 (-TCTT),which accounted for 79.78％(300/376) of total genetic mutations.In addition,a novel β-globin gene mutation CD36 (-C) allele was detected for the first time,the deletion of a nucleotide C at code 36 within exon 2 lead to a frameshift mutation that could result in a premature termination at code 60.Conclusions β-thalassemia mutations in Fujian province are complex with significant genetic heterogeneity.We present for the first time the detection of a new β-thalassemia mutation in the population:CD36(-C),which provides valuable information for genetic counseling and prenatal diagnosis in Fujian province.

OBJECTIVETo investigate the repair of the severe cleft palate in patients over 10 years old.

METHODSFirst, the horizontal palate of the palatine bone was broken and the greater palatine foramen was enlarged by chisel. Then the great palatine neurovascular bundle was released. The soft palate was pushed back and lifted as described by Pro. Ruyao Song. Finally, a buccal musculomucosal flap was transferred to repair the frontal wound after pushing back the soft palate.

RESULTS13 patients aged 10 - 25 years old were treated by this method. All the flaps survived completely. Both the hard and soft palate were lengthened. Velopharyngeal incompetence was corrected very well and the pronunciation improved markedly.

CONCLUSIONSThis method can close the severe cleft palate without tension and lengthen the soft palate. It can correct velopharyngeal incompetence very well and improve pronunciation dramatically. It is especially useful for severe cleft palate in older patients.

Adolescent ; Adult ; Child ; Cleft Palate ; surgery ; Female ; Humans ; Male ; Mouth Mucosa ; transplantation ; Palate, Soft ; surgery ; Surgery, Plastic ; methods ; Surgical Flaps ; Young Adult

OBJECTIVETo observe the effects of fenvalerate (Fen) on ovarian calcium homeostasis.

METHODShGLCs were obtained from pre-ovulatory follicles in an in vitro fertilization program, and were cultured for 72 hours. Changes in cellular [Ca(2+)]i induced by Fen in hGLCs were detected with laser scanning confocal microscopy (LSCM) by using the fluorescent Ca(2+) indicator fluo-3/AM. SD female rats were divided into four groups (control, 1/15LD(50), 1/50 LD(50) and 1/250 LD(50)) in experiment. The activity of ovarian Ca(2+)-ATPase and phosphorylase A (P-a) and the contents of calmodulin (CaM) were assessed after a 30-day Fen exposure. In addition, serum estradiol-17 beta (E(2)) and progesterone (P(0)) concentration were measured by radioimmunoassay, which the sampling rats were ensured at diestrus stage before killed according to vaginal smear.

RESULTS20.0 and 2.0 micromol/L Fen induced the increased of [Ca(2+)]i in hGLC. This [Ca(2+)]i increase mostly resulted from Ca(2+) influx in the studied concentration. Fen had shown the inhibition effects on activity of Ca(2+)-ATPase in 1/250 LD(50) group (P < 0.001) while the activity of phosphorylase A (P-a) in treated groups had significantly enhanced than those of in control. The contents of CaM in ovaries were found to be increased in treated groups. E(2) in 1/250 LD(50) group were higher while P(0) in 1/15 LD(50) group were significantly lower (P < 0.05).

CONCLUSIONExposure to Fen interferes the serum steroid hormone concentrations partly through calcium signal pathway.

Adenosine Triphosphatases ; metabolism ; Animals ; Calcium ; metabolism ; Cells, Cultured ; Female ; Gonadal Steroid Hormones ; blood ; Granulosa Cells ; drug effects ; metabolism ; Homeostasis ; drug effects ; Humans ; Insecticides ; toxicity ; Nitriles ; Ovary ; cytology ; drug effects ; metabolism ; Pyrethrins ; toxicity ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the distribution of deep inferior epigastric perforator vessel and application of the expanded perforator flap for large scar on hand and forearm.

METHODS20 healthy adults were selected to detect the distribution of deep inferior epigastric perforator vessel. 10 cases with cicatricial constriction on hand and forearm were treated with expanded perforator flap.

RESULTS425 perforator points were detected on the skin surface of abdomen in 20 adults. 80% of the points are located within the area which is 1.1 -5.8 cm far from the mid-line of abdomen. According to the Rand method, the percentage of the points in zone I , II , III were 26%, 43%, and 30%, respectively. There were few points in zone IV. The area around navel had a high density of points. The points were distributed asymmetrically at the two sides of abdomen. 10 cases were treated. Vascular deficiency happened at the distal end of one flap. All the other 9 flaps survived.

CONCLUSIONSThe expanded deep inferior epigastric perforator flap is thin and has a large size. It is very suitable for large scar on forearm or hand.

Adolescent ; Adult ; Burns ; complications ; Contracture ; etiology ; surgery ; Epigastric Arteries ; Female ; Humans ; Male ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; blood supply ; Tissue Expansion ; Young Adult

OBJECTIVETo explore the feasibility of IGF2 imprinting system in target gene therapy for tumors.

METHODSThe mouse H19 enhancer, DMD and promoter H19 were amplified by PCR from mouse genomic DNA and then cloned into the plasmid pDC312. The EGFP and DT-A fragments were amplified by PCR and cloned into the recombinant plasmid, and then the shuttle plasmid were transfected into HEK293 cells together with the adenoviral vector Ad5, namely, Ad-EGFP and Ad-DT-A. Adenovirus hexon gene expression was applied to confirm the presence of adenovirus infections. The effect of the IGF2 imprinting system was tested by fluorescence microscopy. RT-PCR and Western blotting after transfection of the recombinant adenoviral vectors into cancer cells were used to show loss of IGF2 imprinting (LOI) and maintenance of IGF2 imprinting (MOI), respectively. The anti-tumor effect was assessed by MTT and flow cytometry after the HCT-8 (LOI). Human breast cancer cell line MCF-7 (MOI) and human normal gastric epithelial GES-1 (MOI) cell line were transfected with Ad-DT-A in vitro. The anti-tumor effect was detected by injecting the Ad-DT-A in nude mice carrying HCT-8 tumors.

RESULTSThe expression of EGFP protein, DT-A mRNA and DT-A protein were seen to be positive only in the HCT-8 tumor cell line. Infection with Ad-DT-A resulted in obviously growth inhibition in HCT-8 cells (75.4 ± 6.4)% compared with that in the control group, and increased the percentage of apoptosis in the HCT-8 cells (20.8 ± 5.9)%. The anti-tumor effect was further confirmed by injecting the recombinant adenoviruses in HCT-8 tumor-bearing nude mice, and the results showed that the Ad-DT-A inhibited the tumor growth, with on inhibition rate of 36.4%.

CONCLUSIONSThe recombinant adenoviruses carrying IGF2 imprinting system and DT-A gene have been successfully constructed, while Ad-DT-A can effectively kill the tumor cells showing loss of IGF2 imprinting. It might play an important role in future target gene therapy against malignant tumors based on loss of IGF2 imprinting.

Adenoviridae ; genetics ; Animals ; Apoptosis ; Breast Neoplasms ; genetics ; pathology ; Colonic Neoplasms ; genetics ; pathology ; therapy ; Diphtheria Toxin ; biosynthesis ; genetics ; Female ; Genetic Therapy ; methods ; Genetic Vectors ; Genomic Imprinting ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; Insulin-Like Growth Factor II ; genetics ; metabolism ; MCF-7 Cells ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Peptide Fragments ; biosynthesis ; genetics ; Plasmids ; RNA, Messenger ; metabolism ; Random Allocation ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

OBJECTIVETo optimize the acupuncture treatment programs for facial paralysis.

METHODSSixty-three patients of facial paralysis were randomly divided and treated according to the table of L9 (3)4 in orthogonal test. They were treated with different combined programs of 4 factors and 3 levels, including factor A (acupuncture op portunity), B (acupoints prescription), C (quantity of stimulus) and D (time of electroacupuncture). The change of facial nerve function score was observed to choose the best acupuncture treatment program for facial paralysis from factor A (acupuncture opportunity), B (acupoints prescription), C (quantity of stimulus), D (time of electroacupuncture) and their 3 levels in each factor.

RESULTSB (acupoints prescription) and D (time of electroacupuncture) were significant factors (P < 0.05), and B (acupoints prescription) was the most important influential factor. B3 (alternative use of two groups of acupoints) was the best one among the 3 levels of B (acupoints prescription), and D3 (electroacupuncture in disperse-dense wave for 30 min) was the best one of D (time of electroacupuncture).

CONCLUSIONTwo groups of acupoints alternatively used with electroacupuncture in disperse-dense wave for 30 min is the best treatment program for facial paralysis.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Facial Paralysis ; therapy ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

OBJECTIVEThis study is to see the pathologic change of cardiac myocyte in Athlete's Heart, and explore the mechanism of the pathologic change.

METHODSFifteen male SD rats were separated randomly into control group (without any exercise), aerobic exercise group (Ae group, swimming for 75 min every day), and overloading exercise group (Oe group, swimming for 180 min with a loading of 5 percent of body weight every day). After 5 days per week for 12 weeks, swimming stopped, the rat hearts were prepared to specimens and examined under Transmission Electron Microscope.

RESULTSThe Ae group, the number and volume of mitochondria increased, and the membrane of mitochondria remained entire. Few of dense bodies were found in cytoplasm. The nucleus envelopes of expansion nucleus appear as dentition. These changes were considered as the adaptation to exercises. At the same time, some pathologic changes of the cardiac myocytes similar to senescence also appeared, such as mitochondria expanse, the crista disorder or disappearance, unclear mitochondria membrane, many dense bodies in cytoplasm, nucleus disfiguration and chromatin collection at edge.

CONCLUSIONAfter exercise training, some pathologic changes of cardiac myocyte also occur with physiological changes. With the raise of exercise intension, the pathologic changes become more obvious, even appearance of cardiac myocyte death.

Animals ; Cardiomegaly ; etiology ; pathology ; Exercise Tolerance ; Male ; Microscopy, Electron ; Mitochondria, Heart ; ultrastructure ; Myocardial Contraction ; Myocytes, Cardiac ; ultrastructure ; Rats ; Rats, Sprague-Dawley ; Swimming