Objective: To explore the feasibility of using siRNA targeting metallothionein 1H (MT1H) gene to reverse the drug resistance of A549/ DDP cells to cisplatin (DDP). Methods: The expressions of MT1H mRNA in A549 and A549/DDP cells were detected by reverse transcriptase polymerase chain reaction (RT-PCR). MT1H siRNA was transfected into A549/DDP cells with high expression of MT1H. The mRNA and protein levels of MT1H were detected by RT-PCR and Dot blotting, respectively. The chemosensitivity of A549/DDP cells to cisplatin was assessed by MMT assay. The apoptotic ratio induced by DDP was determined by TUNEL and flow cytometry analysis (FCM). The apoptosis-related protein levels of Bcl-2 and Bax were determined by immunohistochemistry. Results: MT1H mRNA was highly expressed in A549/DDP cells but not in A549 cells. The mRNA and protein levels of MT1H were significantly down-regulated at 48 h after transfection in MT1H siRNA group compared with the control group. The chemosensitivity of A549/DDP cells to DDP was significantly elevated. The apoptotic ratio induced by DDP was significantly increased. Expression of Bcl-2 was greatly down-regulated but the expression of Bax had little change. Conclusion: Silencing MT1H gene with MT1H siRNA reduces the expression of Bcl-2, enhanced the apoptosis-inducing effect of DDP on A549/DDP cells, and effectively reverses the drug resistance of A549/DDP cells.

Objective To explore the methodology of the preventing the absorption after autog-enous fat granule mammaplasty, and the main point of the operative methods.. Methods The clinical data were reviewed in 30 cases of patients with liposuction combined with fat-grafting injection for breast mammaplasty in the past four years. The tumescent technique was used in liposuction and the volume of fat tissue aspirated was between 500 ml and 3250 ml. Using appropriate way we prepared pure fat granules and then evenly injected into the layer beneath the pectoralis major muscle, submam-mary layer and subcutaneous layer of the breast, respectively. Results The effect of the method was satisfying because sucking area was significantly decreased. From six months to 10 months follow-up, grafts showed no obvious absorption. The breast looked symmetry. Conclusion Fat grafting injection is a practicable method for breast mammaplasty. The key point is to use appropriate way to get pure fat granule and to prevent infection. It deserves to be recommended.

Objective To elucidate whether focal segmental glomerulosclerosis (FSGS) is a secondary development of the COL4-linked thin basement membrane nephropathy (TBMN) or the primary FSGS produces thin glomerular basement membrane (GBM). Methods The family members presented microscopic hematuria,increasing proteinuria with the years and a dual pathological diagnosis of FSGS and TBMN was made in the proband.DNA linkage analysis at locus 2q36-37 that contains the COL4A3/COL4A4 genes was performed with polymorphic micmsateilite markers D2S434,D2S279,D2S1370,D2S256 and D2S427.Haplotypes were constructed at the COL4A3/COL4A4 loci for affected and unaffected family members.All exons of COL4A3 and COL4A4 genes were screened for mutations in the proband.Mutation screening was also performed for NPHS1,NPHS2,CD2AP,WTI,TRPC6 and ACTN4 to exclude familial FSGS.Mutation or polymorphism found in the family were examined in 50 healthy controls. Results In this family hematuria segregated with the 55224 haplotype at the COL4A3/COL4A4 locus.G to A substitution at nucleotide 1214 resulting in an glycine being replaced by glutamate (G405E) was demonstrated for the first time in cxon 20 of COL4A4 gene.G4OSE was present in all four members of the family with hematuria but not in the seven unaffected family members nor in 50 healthy controls.A novel polymorphism segregating with hematuria (IVS1-4C＞T in exon 2 ofCOL4A3) was also found which was only present in all four affected family members but not in the seven unaffected family members. No mutations were demonstrated in FSGS associated genes,however,a novel SNP (R268Q),which distributed with the disease ineompletely,was described in the NPHS1 gene coding nephrin,the podocyte slit diaphragm protein. Conclusions In this family,FSGS occurres on the basis of TBMN.Maybe the particular COL4A3/COL4A4 mutation and polymorphism work together to develop proteinuria and eventually leading to FSGS.But whether the mutation and the polymorphism segregating with the disease predispose to develop FSGS in TBMN patients is required further study.

New field battle X-ray vehicle(model XCY2002-1/200) is used usually and control method is grasped too. By analyzing operation capability, the advantages and disadvantages are found, and then corresponding improvement is proposed to educe efficiency of medical support.

ObjectiveTo investigate the effect of electromyography-triggered neuromuscular stimulation on rehabilitation of hemiplegia patient.Methods20 stroke patients following hemiplegia were treated with electromyography-triggered neuromuscular stimulation on base of routine physical therapy, occupational therapy. Myoelectricity value, active range of motion (AROM) of wrist dorsiflexion, and Fugl-Meyer Assessment(FMA) of upper extremity were assessed before and after the treatment.ResultsThe value of myoelectricity, AROM of wrist dorsiflexion, and FMA of upper extremity have significant statistical improvement (P<0.05) after the treatment.ConclusionElectromyography-triggered neuromuscular stimulation has positive effect on recovery of hemiplegic upper extremity.

OBJECTIVETo discuss whether asiaticosides could effectively reduce the endothelial cell damage as a biochemical modulator, so as to further inhibit the post-stenting intima-media membrane hyperplasia.

METHODHuman aortic smooth muscle cells and aortic fibroblasts were selected and divided into the blank group, the rapamycin group and the asiaticoside group and the rapamycin and asiaticoside group. The expressions of muscle cells and fibroblasts TGF-beta1, Smad7 and I-collagen gene were determined by RT-PCR. The expression quantity of I-collagen protein was assayed by ELISA. The coefficient of drug interaction (CDI) between rapamycin and asiaticoside was calculated. Additionally, 16 Chinese mini-swines were randomly divided into group A and group B. One sirolimus drug-eluting stent of the same type was implanted after the high-pressure pre-expansion of anterior descending artery balloon. After the operation, the group A was intravenously injected with normal saline 30 mL x d(-1). Whereas the group B was intravenously injected with asiaticoside 30 mg x kg(-1) x d(-1)(diluted to 30 mL). The expressions of plasma vWF of the two groups were measured at the 7th and 14th days after the operation. At the 28th day after the operation, tissues of the stented vessel segments were sliced and stained to calculate the vessel area, inner stent area, lumen area and neointima area

RESULTCompared with the control group, the combination group showed significant up-regulation in smooth muscle cells and fibroblast Smad7 gene, down-regulation in TGF-beta, and obvious inhibition of I-collagen gene expression (P < 0.01). As for smooth muscle cells, there was no difference in the expression of I-collagen between the combination group and the rapamycin group, with CDI at 0. 83. As for fibroblasts, there was a significant difference in the expression of I-collagen between the combination group and the rapamycin group (P < 0.05), with CDI at 0.77. Plasma vWF of the group B was significantly lower than that of the group A (P < 0.05) at the 7th and 14th days after the operation. At the 28th day after the operation, no difference was observed in vessel area and stent area between the two groups. However, the lumen area in the group B was significantly larger than that of the group A(P < 0.05), and the neointima area of the group B was significantly smaller than that of the group A (P < 0.05).

CONCLUSIONAs an effective biochemical modulator for rapamycin, asiaticosides could inhibit TGF-beta expression, significantly decrease the synthesis and secretion of extracellular matrix, further inhibit the post-stenting intima-media membrane hyperplasia and reduce the endothelial cell damage by effectively up-regulate the expression of Smad7 protein.

Animals ; Collagen ; genetics ; metabolism ; Coronary Restenosis ; drug therapy ; prevention & control ; surgery ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Hyperplasia ; drug therapy ; genetics ; metabolism ; prevention & control ; Smad7 Protein ; genetics ; metabolism ; Stents ; adverse effects ; Swine ; Transforming Growth Factor beta1 ; genetics ; metabolism ; Triterpenes ; administration & dosage

OBJECTIVETo test the bovine cementoblasts (CBs) cementum-forming ability in vivo.

METHODSRoot fragments of newborn bovine freshly extracted mandibular incisor were cultured routinely and 4th-5th passages of CBs were harvested. CBs were then cultured in the medium supplemented with 50 mg/L alpha-ascorbic acid and 10 mmol/l beta-glycerolphosphate to form a thick layer as tissue engineering scaffold for cementum formation. Collagen membrane was used as control scaffold. 2 x 10(6) cells were attached to the CBs-made carrier as well as collagen membrane scaffolds and transplanted subcutaneously into immunodeficient mice. Transplants were harvested at 7th week. Histological sections were stained with HE, alizarin red S and van Kossa methods as well as monoclonal Ab against bovine cementum attachment protein (CAP).

RESULTSCBs-made scaffold supported more cementum-like tissue (CLT) formation than collagen-made scaffold. The CLT formed on CBs scaffold was partly calcified with embedded cells. Uncalcified cementoid-like material could be seen on the surface and was encircled by cubical CB-like cells. The CLT was also positive to CAP and van Kossa staining.

CONCLUSIONSThese results suggest that the bovine CBs can form cementum-like tissue. The cell-made carrier is a better scaffold than collagen membrane.

Alkaline Phosphatase ; analysis ; Animals ; Bone Transplantation ; methods ; Cattle ; Cell Adhesion Molecules ; analysis ; Cells, Cultured ; Dental Cementum ; chemistry ; cytology ; transplantation ; Immunohistochemistry ; Integrin-Binding Sialoprotein ; Male ; Mice ; Mice, Nude ; Osteocalcin ; analysis ; Osteonectin ; analysis ; Sialoglycoproteins ; analysis ; Tissue Engineering ; methods ; Transplantation, Heterologous

BACKGROUNDRetroviral vectors have been widely used to introduce foreign into various target cells in vitro, thus showing relatively high systemic delivery efficiency of various transgene products. The authors investigated the stability and efficiency of skeletal muscle-specific hybrid retroviral vectors in expression of human factor IX (FIX) in vitro and iv vivo.

METHODSFIX cDNA in LIXSN vector was replaced with a FIX minigene containing splicing donor and splicing acceptor sequence of first intron of human FIX gene. Two copies of muscle creatine kinase enhancer (MCK, Me2) were inserted in forward or reverse orientation at NheI site of 3' long terminal repeat (LTR), resulting in two hybrid vectors, which were designated as LMe2IXm2SN(F) and LMe2IXm2SN(R), respectively. The vectors were tested in vitro and in vivo for stability and muscle-specificity of factor IX expression with SCID mice.

RESULTSMuscle cells carrying vector with Me2 expressed significantly higher levels of FIX (up to 1800 ng/106.24 h) than those without Me2, thus suggesting that Me2 could specifically increase expression level of FIX in muscle cells. Myoblasts transduced with LMe2IXm2SN(R) produced much less FIX in vivo in SCID mice than LMe2IXm2SN(F). One or two copies of Me2 sequence were deleted in myoblasts transduced with LMe2IXm2SN(R) without changing the orientation of Me2.

CONCLUSIONSLTR inserted with MCK enhancers can specifically increase human FIX expression in skeletal muscle cells in vitro and in vivo, and MCK enhancer should be positioned in the same orientation as that of LTR promoter.

Animals ; Creatine Kinase ; genetics ; Creatine Kinase, MM Form ; Enhancer Elements, Genetic ; Factor IX ; analysis ; genetics ; Gene Expression ; physiology ; Genetic Techniques ; Genetic Vectors ; Hybridization, Genetic ; Isoenzymes ; genetics ; Mice ; Mice, SCID ; Retroviridae ; genetics ; Terminal Repeat Sequences

Objective To evaluate the value of 3.0T magnetic resonance multi-b value diffusion-weighted imaging (DWI) in evaluating the efficacy of chemotherapy for patients with central lung squamous cell carcinoma and atelectasis. Methods Twenty patients with lung squamous cell carcinoma were examined by magnetic resonance imaging (MRI) (including T1WI, T2WI and multi-b value DWI) before chemotherapy, 2 cycles of chemotherapy and 4 cycles of chemotherapy. The images, the tumor volume and changes of apparent diffusion coefficient (ADC) were analyzed. Results In the patients with central lung cancer and atelectasis, the tumor and atelectasis could be distinguished on MRI examination before radiotherapy. It was more easily identified on T2WI images after radiotherapy. In the 20 patients, the ADC values in the effective group (partial remission or complete remission) and the invalid group were increased, but the differences of ADC values in the effective group before chemotherapy, 2 cycles and 4 cycles of chemotherapy were statistically significant [b=800 s/mm2:(1.09 ± 0.52) × 10-6 mm2/s, (1.22 ± 0.59) × 10-6 mm2/s, (1.24 ± 0.52) × 10-6 mm2/s, F = 31.19, P < 0.001]. There was no significant difference in ADC values between before and after chemotherapy (b = 800 s/mm2: (1.10 ± 0.49) × 10-6 mm2/s, (1.16 ± 0.60) × 10-6 mm2/s, (1.20 ± 0.72) × 10-6 mm2/s, F=2.86, P=0.089]. When b=800 s/mm2, the ADC curve slope in the effective group was more stable, better linearity. Conclusions The MRI technique can accurately distinguish the tumor from atelectasis before and after chemotherapy. The change of ADC value after chemotherapy is earlier than that of morphological change. The change rate of b value can better evaluate the curative effect of chemotherapy.

OBJECTIVETo develop and evaluate the short version of patient reported outcomes (PROs) questionnaire for gastric stuffiness (Wei Pi) patients with modern test theory and technologies, hoping to provide testing tools for related clinical practice and scientific researches with higher quality and less administrative and response burdens.

METHODSUsing descriptive study design, clinical data were collected with sociological questionnaire and previous developed full items version of PROs instrument for gastric stuffiness (Wei Pi) patients via field and online surveys between Sep 2011 and Mar 2012. The statistical analysis group identified the termination parameters firstly, and then selected items with discrimination, fitting residual, item information curve (IIC) , item characteristic curve (ICC), and the rank of computerized adaptive testing (CAT) select proportion, etc. After assumption evaluation of item response theory (IRT), IIC, ICC, difficulty coefficient distribution, items-response relation and thresholds, etc. were used for psychometric evaluation of instrument.

RESULTSA total of 331 patients [Ages: 31.99 +/- 10.29 yrs; Male: 186 (56.3%)] were enrolled in statistical analysis. The test termination criterion was Max SE = 0.2 or Max items number =16. After items selection, a 15-item short version of instrument, which contains symptoms facet (8 items) and impact facet (7 items) was generated. With good unidimensionality, local independence, and monotonicity, the IC and ICC in IRT analysis showed good working capability of the questionnaire. The difficulty coefficient distribution and items-response relation were also rational, as well as response thresholds.

CONCLUSIONSThe short version of PROs instrument for adult gastric stuffiness (Wei Pi) patients was successfully developed and assessed. The instrument with good methodological and reporting quality could be used in clinical and scientific evaluating their symptoms and impact.

Adult ; Computer Simulation ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Psychometrics ; Stomach Diseases ; diagnosis ; therapy ; Surveys and Questionnaires ; Young Adult