Objective: To compare the efficacy of moxibustion with different doses for knee osteoarthritis (KOA), and explore the correlation between moxibustion dose and clinical efficacy. Methods: Sixty-eight patients with KOA who met the inclusion criteria were randomly divided into a 20-minute moxibustion group and a 40-minute moxibustion group by the random number table method, with 34 cases in each group. Dubi (ST 35), Neixiyan (EX-LE 4) and Heding (EX-LE 2) were used for moxibustion in the two groups. Each treatment lasted 20 min or 40 min for each point in the 20-minute moxibustion group and 40-minute moxibustion group, separately; the treatment was given 3 times a week and lasted for 4 weeks. The visual analog scale (VAS), Western Ontario and McMaster University osteoarthritis index (WOMAC) and traditional Chinese medicine (TCM) symptom scores were evaluated before and after treatment to compare the efficacy between different moxibustion doses for KOA. Results: After treatment, the total effective rate was 87.5％ in the 40-minute moxibustion group, versus 70.0％ in the 20-minute moxibustion group, and the difference in the total effective rate between the two groups was statistically significant (P<0.05). After treatment, the VAS scores, the total WOMAC scores and the component scores of pain, stiffness and dysfunction, and the TCM symptom scores in both groups all changed significantly when compared with those before treatment (all P<0.05). After treatment, the between-group differences in the VAS score, the total WOMAC score and the component scores of pain and dysfunction, and the TCM symptom score were statistically significant (all P<0.05), while the difference in the stiffness score in WOMAC showed no statistical significance (P>0.05). Conclusion: Either 20-minute moxibustion or 40-minute moxibustion can relieve pain, improve stiffness, dysfunction, and TCM symptoms for KOA; and 40-minute moxibustion is better in relieving pain, improving dysfunction and TCM symptoms.

Objective To construct eukaryotic expression plasmid pIRESneo3-pigment epithelium-derived factor (PEDF) and detect its transient expression in SP2/0 cells. Methods Specific primers were designed based on the mature peptide sequence of human PEDF cDNA in the GenBank. Human PEDF gene was cloned into the eukaryotic expression vector pIRESneo3. The PEDF DNA was transfected into SP2/0 with LipofectamineTM 2000. The recombinant human PEDF protein expressed in SP2/0 cell culture supernatant was identified by Western blot and enzyme-linked immunosorbent assay. The biological activity of the recombinant human PEDF was measured by 3-(4,5-dimethylthiazol-z-y1)-2,5-diphenytetrazolium bromide(MTT) method. Results PCR amplification, restriction enzyme digestion and DNA sequencing confirmed that the mature peptide sequence of human PEDF cDNA was successfully cloned into the eukaryotic expression vector pIRESneo3. And the plasmid was transfected into SP2/0 cells, which could secret PEDF. Western blot analysis showed that there was only one obvious band at the position of relative molecular weight of 50 000, and it is equivalent to the expected value. Enzyme-linked immunosorbent assay suggested that the content of PEDF began to rise after transfection, and peaked at 36 h [(0.92±0.04) μg/ml]. The proliferation of human umbilical vein endothelial cell line was significantly inhibited by supernatant after transfection of 36 h (P<0.05). Conclusions The eukaryotic expression plasmid pIRESneo3-PEDF had been successfully constructed and active human PEDF was transiently secreted, which made a foundation for further study of stable expression and purification of PEDF. This protein could be a potential medication for preventing and managing retinopathy of prematurity.

Objective　To examine whether ischemia preconditioning (IPC) can decrease ischemia reperfusion(I/R) injury in retina and investigate the mechanism. Methods　Retinal ischemia was induced in SD rats by increasing the intraocular pressure to 14.63kPa for 60 minutes via cannulation into the anterior chamber. Retinal ischemia for 5 minutes constituted the preconditioning stimulus. Rats first underwent IPC and 60 minutes of ische- mia 30 minutes, 24 or 72 hours later or they underwent a 5-minute sham PC and 60 minutes of ischemia 24 hours later. Their electroretinography(ERG), Light microscopy(LM),transmission electron microscopy (EM) and the expression of heat shock protein 70(HSP70) were observed and the content of malandialdehyde(MDA) in rat retinas were measured 24 or 72 hours later after 60 minutes ischemia.Results　Ischemia induced histologic damage was completely prevented when IPC was performed 24 or 72 hours (but not 30 minutes) before ischemia. In contrast to the sham PC group, the amplitudes had complete recovery of b-waves with preischemia basline amplitudes (P<0.01),and the content of MDA was significantly decreased (P<0.01).Conclusion　IPC provides protection against retinal I/R injury just during limited period of time.

Objective To evaluate surgical treatment of infected femoral artery pseudoaneurysm.Methods The data on surgical treatment of 45 patients with infected femoral artery pseudoaneurysm admitted from January 2003 to June 2008 were analyzed retrospectively.Fourty-three patients underwent operative treatment including excision of infected femoral artery pseudoaneurysm,exhaustive debridement and bypass graft with vascular prosthesis.Two patients were unavoidable to undergo removing of infected femoral artery pseudoaneurysm and ligating the proximal and distal artery of pseudoaneurysm because of severe infection and large volume.Results The patients were followed up from 3 to 12 months(mean 7.82 months).The limbs of all the patients underwent bypass graft with vascular prosthesis were salvaged successfully,patients of which had secondary wound healing and had not intermittent lameness.One of two patients performed ligation of artery was salvaged successfully but had severe intermittent lameness,another patient underwent high amputation above knee because of ischemic gangrene.ConclusionFor infected femoral artery pseudoaneurysm,the operative treatment including excision of infected femoral artery pseudoaneurysm,exhaustive debridement and bypass graft with vascular prosthesis is effective and safe.

AIM: To provide experimental evidence for gene therapy of thrombophilia disease, we constructed the eukaryotic expression plasmid with human thrombomodulin (hTM) gene and observed the alteration of hTM expression on the surface of human umbilical vein endothelial cells (HUVECs) with and without the reconstructive plasmid. METHODS: The whole expressive fragment of hTM gene was amplified by PCR from human genome. Both hTM gene and pcDNA3.1(+)/neo empty vector was digested by HindⅢ and EcoRⅠ. Two digested fragments were ligated into pcDNA3.1/hTM with T_4DNA ligase. After identifying, the reconstructive plasmid transfected into HUVECs using lipofectin. The hTM antigen on the HUVECs was detected by immunohistochemistry. RESULTS: The hTM reconstructive plasmid was confirmed by double endonuclease redigesting and sequencing. About 10% HUVECs were transfected by pcDNA3.1/hTM plasmid with lipofectin and the high-level hTM was detected on the transfected cells. CONCLUSION: We constructed the pcDNA3.1/hTM plasmid successfully, and it could be expressed on the HUVECs. [

Glucocorticoids (GC) exert a broad effect on the body and have been extensively used clinically.It is well known now that GC can act via both genomic and nongenomic mechanisms.Genomic effects of GC have been well reviewed elsewhere.Here we focus on the current understanding of nongenomic effects and discuss their biologic significance as well as the remaining problems in this field.

Objective To summarize the clinical presentations, the findings of lab tests and procedures and the genetic investigation of collagen type Ⅵ related myopathy, and to help clinicians recognize and diagnose this rare disease.Methods Seven familiar or spontaneous collagen type Ⅵ related myopathy patients diagnosed by gene detection were analyzed.We emphasized on the features of clinical manifestations, serum creatine kinase level, electromyography, lower-limb muscle MRI, muscle biopsy and correlation between genotype and pZenotype.Results Among 7 patients, 3 were caused by COL6A1 mutation, 1 was caused by COL6A2 mutation and 3 were caused by COL6A3 mutation.Two patients were familiar wZile 5 were spontaneous.HigZligZted clinical presentations were proximal weakness in lower limbs and joint contrature.Serum creatine kinase level was sligZtly elevated.ElectromyograpZy sZowed sligZt myogenic damage.Muscle MRI of tZigZ sZowed distinct pattern of muscle involvement.Muscle patZology revealed dystropZic myogenic cZanges with proliferation of connective tissue between muscle fibers.Conclusions Neurologists should recognize the features of collagen type Ⅵ related myopathy, such as progressive weakness, early-onset joint contraetures, slightly elevated serum creatine kinase and selective muscle involvement on leg MRI scan, and then perform next-generation sequencing based genetic test on suspected patients.This approach would improve the diagnostic rate of the disease.

Objective:To observe the clinical effect of mild moxibustion for knee osteoarthritis (KOA) and to explore the role of mild moxibustion in relieving pain,reducing stiffness and improving joint dysfunction in patients with KOA.Methods:Eighty patients with KOA were randomly allocated into either a moxibustion group or a medication group by the random number table,with 40 cases in each group.The moxibustion group used mild moxibustion at Neixiyan (EX-LE 5),Dubi (ST 35),Xuehai (SP 10) and Liangqiu (ST 34),30 min each time,3 times a week;the medication group was given celecoxib capsule (celebrex),0.2 g each time,once a day.Both groups were treated for 4 weeks.The visual analog scale (VAS) and Western Ontario and McMaster Universities osteoarthritis index (WOMAC)scores were evaluated before and after treatment.The efficacy of the two groups was compared after treatment.Results:After treatment,the overall efficacy of the moxibustion group was significantly different from that of the medication group (P＜0.05).The VAS and WOMAC scores of the two groups were lower than those before treatment (both P＜0.01).The changes in the VAS and WOMAC scores after treatment in the moxibustion group were significantly different from those in the medication group (both P＜0.05).After treatment,in single item of WOMAC,the changes in pain and joint dysfunction in the moxibustion group were more statistically significant than those in the medication group (both P＜0.05).Conclusion:Mild moxibustion and oral celebrex can reduce the VAS and WOMAC scores of patients with KOA.Mild moxibustion is superior to oral celebrex in relieving pain and improving joint function.

Objective To explore the effection of four types of surgery for the treatment of adenoid hypertrophy . Methods 148 cases of with adenoid hypertrophy treated in our hospital between April 2012 and April 2015 were chose;they were randomly divided into 4 groups ,each group of 37 people .A group of patients with adenoid hypertrophy were taken traditional ade‐noidectomy curettage;Group B with nasal endoscopic adenoidectomy and cutting aspiration biopsy ;Group C by adenoidectomy shave their + residual endoscopic adenoidectomy bit cut method combined treatment ;Group D with nasal endoscopic adenoidectomy plas‐ma cutting treatment .The curative effect ,operation time ,blood loss were observed ;patients were followed‐up for half a year ,ade‐noidectomy residual rate and complications of each group were compared .Results The total effective rate of B ,C ,D three groups were significantly higher in group A patients (χ2 =7 .731 ,5 .045 ,7 .731 ,P<0 .05) ,the efficient between three groups was not sta‐tistically different (P>0 .05) .B ,C ,D three groups of operation time is significantly higher than A group of patients (t=5 .819 , 5 .829 ,2 .759 ,P<0 .05);B and C group had long operation time than group D (t=3 .555 ,3 .637 ,P<0 .05);But operation time of B and C had no significant difference between the two groups (t=0 .149 ,P>0 .149) .Bleeding of B and C group were significantly higher than group A (t=9 .305 ,4 .126 ,P<0 .05);Group D was significantly lower than A ,B ,C three group (t=8 .054 ,16 .559 , 12 .837 ,P<0 .05);Group C and group B was significantly higher than the bleeding (t=5 .739 ,P<0 .05) .Retention rate of group A is significantly higher than the other three groups (χ2 =31 .308 ,31 .308 ,24 .667 ,P<0 .05) ,the residual rate of B ,C ,B group were lower ,there was no statistically significant difference(P> 0 .05) .Complication rates between the four groups was no statistical difference(P>0 .05) .Conclusion we should choose the right means of surgical treatment according to patients condition and eco‐nomic situation to .

Background The multipotent differentiation features of induced pluripotent stem cells (iPSCs) offer a new option for cell replacement therapy of many clinical diseases.In ophthalmology,iPSCs are a good model in studying the pathogenic mechanism of degenerative ocular diseases.A better identification method for iPSCs is critical for analyzing the in vivo biological characteristics of iPSCs.Objective This study was to investigate the feasibility and stability of labeling iPSCs with quantum dots.Methods Human umbilical mesenchymal stromal cells-iPSC lines were cultured and amplified on matrigel,and the characteristics of iPSCs were evaluated by immunofluorescence.Different concentrations (5.0,7.5 and 10.0 nmol/L) of quantum dots with a CdSe/ZnS nuclear shell structure were used to label iPSCs after passaging and proliferation.The labeling outcome was observed with a three-dimensional deconvolution real-time live cells imaging system.The labeled iPSCs were subsequently cultivated,and then changes in fluorescence intensity were examined 7 days after the first and the second passaging of iPSCs.Results iPSCs were observed to grow in a clonal manner under the inverted microscope.The iPSC markers,OCT4 and Nanog,were detected by immunofluorescence.With increasing concentrations of quantum dots,the fluorescence intensities representing the levels of OCT4 and Nanog in iPSCs were gradually elevated,with optimal levels of fluorescence observed at a concentration of 10 nmol/L of quantum dots.The fluorescent labeling of OCT4 and Nanog in iPSCs remained and weakened gradually till day 7 even after the second passage.Conclusions Quantum dots labeling could be used to track iPSCs in a dose-independent manner.The fluorescent signal from the quantum dots labeling the iPSCs lasts 2 weeks at least.