Sixty-four cases of plantalgia were treated by warm needling at Yongquan ( KI 1 ), Lineiting ( Ex-LE) and an empirical point. Thirty cases were cured, 28cases improved and 6 cases failed.

BACKGROUND: No particular marker molecule of bone marrow mesenchymal stem cells (BMSCs) is presently found, so its determination is difficult. BrdU marker has no radioactive pollution. Some studies have confirmed that BrdU marker has no damage to cell function without ultraviolet radiation. OBJECTIVE: To investigate in vitro identification and labeling methods and biological characteristics of adult BMSCs. DESIGN, TIME AND SETTING: The cell experiment was conducted at the General Hospital of Lanzhou Military Area Command of Chinese PLA between June and December 2007. MATERIALS: Bone marrow was collected from 5 healthy adult volunteers. BrdU was purchased from Sigma, USA. METHODS: 10 mL adult bone marrow was harvested to isolate mononuclear cells by density gradient. Cells were cultured in DMEM containing 10% fetal bovine serum and proliferated at a density of 2?108 L-1. At the third passage, BMSCs was inoculated in medium supplemented with osteoblast inductor and stained with alkaline phosphatase. Subsequently, BMSCs were inoculated in medium supplemented with lipoblast inductor and stained with oil red O. Cells were incubated with BrdU at different concentrations of 5, 10 and 15 ?mol/L for 12, 24, 48, 72 and 96 hours, and then detected by immunocytochemistry. MAIN OUTCOME MEASURES: Cell growth and proliferation were observed under an inverted light microscope. Cell phenotype, osteoblast and lipoblast differentiation were identified by flow cytometry. BrdU-positive labeling rate and cell growth after labeling were investigated. RESULTS: In vitro cultured BMSCs were homogenous population and exhibited a spindle-shaped fibroblastic morphology. BMSCs expressed CD44 and CD71, but did not express CD34 and HLA-DR. BMSCs can differentiate into osteoblasts and lipoblasts. Most BMSCs were labeled by BrdU. With the increase in labeling concentration and over time, BrdU positive rate gradually increased and exceeded 90% after labeling with BrdU (10 ?mol/L) for 72 hours, with the labeling identifiable in nine consecutive passages. At the third passage, 90% BMSCs were in G0/G1 phase, and 88.62% in G0/G1 phase after labeling. BrdU has little effect on morphous and proliferation of labeled cells. CONCLUSION: Adult BMSCs can be identified through morphological character, specific surface antigens and multipotential differentiation in vitro. BrdU labeling provides a feasible means for a dynamic observation of the survival, growth and differentiation of the implanted BMSCs. The optimal dosage and timing of BrdU labeling are respectively 10 ?mol/L and 72 hours.

Objective To investigate the expression of Toll-like receptor 2 and 4 (TLR2 and TLR4) in peripheral blood mononuclear cells (PBMCs) and the serum inflammatory cytokine levels in patients with liver failure. Methods The expressions of TLR2 mRNA and TLR4 mRNA in PBMCs were detected by RT-PCR in 20 healthy controls, 20 chronic hepatitis B (CHB) patients, 18 liver failure patients in early stage and 14 in intermediate-end stage. The serum contents of endotoxin, TNF-α and IL-6 were detected by ELISA. Results Compared with the healthy controls, the expression of TLR2 mRNA and TLR4 mRNA, and the contents of endotoxin, TNF-α and IL-6 increased in CHB patients and liver failure patients ( both early stage and intermediate-end stage) ( F = 32.997, 37.476, 23. 951,57. 265 and 38. 403, P < 0.01 ). TLR2 mRNA expression in liver failure patients in intermediate-end stage was higher than that in the early stage, but that for TLR4 mRNA was lower than that in early stage. The expressions of TLR2 mRNA and TLR4 mRNA in PBMCs were significantly correlated with the contents of TNF-α and IL-6 ( r = 0. 917 and 0. 788, P < 0. 01 ). Conclusion The inflammation reaction mediated by TLR2 and TLR4 might participate in the pathogenesis of liver failure.

Objective To investigate the effect and mechanism of adrenomedullin (AM) on apoptosis of renal tubular epithelial cell in rats induced by renal ischemia reperfusion injury. Methods Thirty-two Wistar rats were randomly divided into 4 groups: control group, IRI group, empty plasmid group and AM group. One week after removing the right kidney, eukaryotic expression vector encoding rat AM gene was transfected into the left kidney using an ultrasound-microbubble mediated system. After 1 week the transfer efficiency was detected by immunohistochemical method . Renal IRI model induced by clamping left renal arteries for 45 min followed by reperfusion for 24 h. Tubular cell apoptosis was detected by TUNEL assay. Bcl-2, Bax and Fas expressions were examined by RT-PCR. The expressions of caspase-3, caspase-8 and caspase-9 were determined by Western bolt analysis. Results The expression of AM in the AM group was significantly higher than the empty plasmid group (0.51±0.09 vs 0.23±0.05; P<0.05). Compared with the control group, the apoptosis rate of renal tubular cell in the IRI group was significantly higher [(38.79±7.52)% vs (2.89±0.52)%; P<0.05]. The expressions of Bax, Bcl-2, Fas, caspase-3, caspase-8 and caspase-9 were also significantly increased (0.72±0.18 vs 0.23±0.04, 0.80±0.12 vs 0.38±0.06, 1.24±0.25 vs 0.39±0.09, 0.76±0.13 vs 0.38±0.08, 0.92±0.14 vs 0.32±0.06, 0.89±0.12 vs 0.42±0.09; P<0.05). Bax/Bcl-2 was also significantly increased (0.91±0.18 vs 0.61±0.08; P<0.05). Compared with the IRI group, AM pretreatment significantly decreased the apoptosis rate of renal tubular cells [(19.36±6.78)% vs (38.79±7.52)%; P<0.05]. AM inhibited the up-regulation of Bax, Fas, caspase-3, caspase-8 and caspase-9, while promoting the up-regulation of Bcl-2 (0.48±0.11 vs 0.72±0.18, 0.62±0.07 vs 1.24±0.25, 0.53±0.08 vs 0.76±0.13, 0.46±0.08 vs 0.92±0.14, 0.51±0.12 vs 0.89±0.12, 1.23±0.25 vs 0.80±0.12; P<0.05). Bax/Bcl-2 significantly decreased (0.44±0.12 vs 0.91±0.18; P<0.05). The above parameters had no significant diffe-rence between the empty plasmid group and the IRI group (P>0.05). Conclusion AM can reduce apoptosis of renal tubular epithelial cell induced by renal IRI, the mechanism of which might be achieved by inhibiting caspase-dependent intrinsic and extrinsic pathways.

Risk management penetrate the entire process of medical device regulation, and it is also very necessary for medical devices in use. Based on the analyzing of the status of risk management for medical devices, this paper discusses the principal, participants and entry point of risk management for medical devices.
Device Approval ; Equipment Safety ; Equipment and Supplies ; Risk Management

Objective To investigate the relationship between the homocysteine,sperm DNA fragmentation index and sperm counts of male with severe impaired spermatoqenesis.Methods From December 2015 to February 2017,56 male patients with severe impaired spermatoqenesis were enrolled in the study.The patients were divided into two groups according to the WHO criteria:severe oligozoospermia and azoospermia group (n =25) and oligoasthenoteratozoospermia group (n =31),and the control group was a male with no reproductive impairment (n=27).The sperm parameters were analyzed by using the computer automatic semen analyzer,sperm DNA fragmentation index and serum Hcy level were detected by sperm chromatin diffusion method and enzyme colorimetric method.Results The median of Sperm DNA fragmentation index and homocysteine level in control groups were 33％ [95％CI(29.0％ ～34.4％)] and 13.2 μmol/L [95％CI(12.4 μmol/L～14.2 μmol/L)],and in severe spermatogenesis groups in these two indicators were 21％ [95％CI(19.0％ ～24.0％)] and 8.9 mol/L [95％CI(8.4 μmol/L～ 9.4 μmol/L)],respectively.The results of these two items were higher than the control group,the difference was statistically significant (t=6.793～7.543,P=0.000).Sperm survival rate in normal control group and severe spermatogenesis group was 71％ [95％ CI(67.8％ ～75.1％)] and 57％[95％CI(52.3％ ～58.0％)],respectively,and the difference was statistically significant (t=-8.475,P=0.000).Sperm DNA fragmentation index was positively correlated with serum Hcy level and sperm concentration,Passing-Bablok regression analysis was:Y=10.705 +0.053X,Y=21.071+0.286X,and Hcy level was negatively correlated with sperm concentration.Conclusion The increase of Hcy level and sperm DNA fragmentation index may be an importantcause of male with severe impaired spermatoqenesis,but the specific mechanism remains to be further studied.

Objective To investigate the effect of intermedin (IMD) on the expressions of angiogenesis-related genes induced by renal ischemia reperfusion injury (IRI).Methods Wistar rats were randomly divided into four groups: control group,IRI group,empty plasmid group and IMD plasmid group.One week after removing the right kidney,eukaryotic expression vector encoding rat IMD gene was transfected into the left kidney using an ultrasound-microbubble mediated system.Renal IRI model was induced by clamping left renal arteries for 45 minutes followed by reperfusion for 1 d,2 d,3 d,4 d,7 d and 14 d.The expressions of hypoxia inducible factor-1α (HIF-1o),vascular endothelial growth factor (VEGF) and angiopoietin receptor Tie-2 were examined by RT-PCR and Western boltting.Results Compared with control group,an increase in HIF-1α,VEGF and Tie-2 was observed in the IRI group at d 1,d 2 and d 3 (allP＜0.05).The expression of HIF-1o peaked at d 1 d(P＜0.05),while VEGF and Tie-2 at d 2 (P＜0.05),followed by a decrease that was similar to the control levels at d 4(P＞0.05).Compared with the IRI group,the expressions of HIF-1α,VEGF and Tie-2 of IMD group were much higher and all reached the peak at d 1 (P＜0.05),maintained at d 2-4 (P＜0.05),followed by a decrease at d 7(P＞0.05).The above indexes had no differences between empty plasmid group and IRI group(P＞0.05).Conclusions IMD pretreatment may play an important role in the process of repair and regeneration after renal ischemia reperfusion injury byimproving the expressions of angiogenesis-related genes(HIF-1α,VEGF and Tie-2) induced by renal ischemia reperfusion injury.

Objective To investigate the effects of intermedin (IMD) pretreatment on associated repairing genes of the rats with injured kidneys by ischemia reperfusion injury (IRI)during the repair and regeneration process.Methods A total of 144 healthy male Wistar rats were randomly divided into 4 gourps:sham opration group,IRI group,empty plasmid group (EP)and IMD group.After resection of right kidneys of the rats,plasmid was transfected into the left kidneys by using ultrasonic microbubble technology.After one week,the renal IRI models were programmed.The samples of renal tissues after 1 d,2 d,3 d,4 d,7 d and 14 d of reperfusion were harvested respectively,then the mRNA expressions of the Pax-2,ZO-1,Ncam,Wt-1 and vimentin in renal tissues were detected by RT-PCR; the protein expressions of Pax-2,Wt-1 and vimentin were analyzed by Western blotting and the protein expressions of ZO-1 and Ncam were measured by ELISIA.Results (1) Compared with the sham group,the mRNA and protein expression of Pax-2,ZO-1,Ncam,Wt-1 and vimentin of the rats in IRI group increased significantly from day 1 to day 3 (P＜0.05),which peaked at day 2.After day 4,the above expressions in IRI group returned to normal level.(2) In IMD group,the mRNA and protein expressions of Pax-2,Zo-1,Ncam,Wt-1 and vimentin were significantly higher than those in other 3 groups (P＜0.05) at the same time after IRI day 1 to day 4,with a maximum in day 2.(3) The above expressions in IRI group,EP group,IMD group had no significant differences compared with sham group after day 7 or day 14 respectively (P＞0.05),and either between IRI group and EP group,though the expressions of the genes in IRI group and EP group increased compared with sham group after day 4.(4) The above expressions between IRI group and EP group also had no significant difference (P＞0.05).(5) Changes of ZO-1 and Ncam protein expression detected by ELISA were similar to those abore mRNA and protein expression by RT-PCR and Western blotting.Conclusion IMD pretreatment plays an important role in up-regulation of the expressions of associated repair genes during the process of repair and regeneration after renal ischemia reperfusion injury.

Objective To investigate clinical characteristics of elderly patients with sepsis combined with congestive heart failure and risk factors for short-term mortality.Methods Clinical data of elderly patients with sepsis combined with congestive heart failure who were admitted in our hospital from January 2013 to January 2018 were selected and retrospectively analyzed.They were divided into the survival group(n=134)and the death group(n=83)according to survival status during hospitalization.The clinical characteristics and risk factors for mortality were analyzed and compared.Results A total of 217 elderly patients were enrolled,with 113 males and a mean age of(72.3 ± 7.5)years.The death rate of sepsis was 38.3％ (83/217 cases),and 29 cases died of sepsis and 54 cases died of other diseases.Pneumonia accounted for 78.8％ (171/217 patients) in all patients of two groups,and skin and soft tissue infection for 12.9 ％ (28/217 cases).There were significant differences between two groups in age,body mass index,smoking,diabetes,chronic obstructive pulmonary disease,mean arterial pressure,arterial oxygen partial pressure(PaO2),C-reactive protein,white blood cell counts,neutrophil and lymphocyte counts,glomerular filtration rate,serum sodium level,albumin level,lactate level,and left ventricular ejection fraction(P ＜0.05).Furthermore,the rates of invasive mechanical ventilation and continuous renal replacement therapy were higher in the death group than in the survival group(x2=13.209 and 7.402,P＜0.001 and 0.007).Multivariate Cox regression analysis showed that advanced age,chronic obstructive pulmonary disease,low albumin level and low glomerular filtration rate were risk factors for mortality(P＜0.05).Conclusions Elderly patients with sepsis combined with congestive heart failure often have severe pneumonia and violent skin and soft tissue infection,with worse heart and renal function.Advanced age,chronic obstructive pulmonary disease,low albumin level and low glomerular filtration rate are risk factors for mortality.