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BACKGROUND:Human skin-derived precursors can be cultured for a long term in vitro, and differentiated into neurons, glial cel s, smooth muscle cel s, Schwann cel s and cel s with peripheral neurons phenotype.
OBJECTIVE:To investigate the culture conditions and multiple differentiation capacity of multipotential stem cel s from human skin, especial y the potentials of differentiating into neurons and osteoblasts.
METHODS:Human skin-derived precursor cel s were cultured with trypsin digestion method, and identified with immunocytochemistry. Cel s at passages 3-4 were induced to differentiate into neurons and osteoblasts, and underwent von Kossa staining protocol for calcium, chondrocyte induction, toluidine blue staining, immunohistochemical staining and Sudan black staining. The expression of nestin, vimentin,βIII-tubulin, S100 and col agen II in the human skin-derived precursors was detected.
RESULTS AND CONCLUSION:The human skin-derived precursor cel s cultured with trypsin digestion method could proliferate and form suspending spheres, and nestin positive cel s were detected at any time point of the culture. Al the cultured cel s expressed vimentin, and some adherent cel s expressedβIII-tubulin. Human skin-derived precursor cel s were induced with Salvia miltiorrhiza to differentiate into neuron-like cel s, and expressed marker of nerve cel s. Skin-derived precursors could be induced to differentiate into osteoblasts and von Kossa staining displayed black calcified nodules in the culture dish. Skin-derived precursors could also be induced to differentiate into chondrocytes, and toluidine blue staining was strongly positive, and some cel s expressed col agen II, which suggested that, the differentiated cel s contained chondrocytes. Experimental findings indicate that, skin contains multipotential stem cel s that are capable of differentiating into osteoblasts, chondrocytes, Schwann cel s and oligodendrocytes.

The susceptibility to 9 kinds of antibiotics of 393 animal pathogenic E. coli isolated from clinical samples was determined from 2000 to 2003. The resistance to TC, GM, CMP, AMP, RA, KM, FT, SM and CIP were 93.89%, 57.76%,78.63%, 77.86%, 92.11% ,47.33%, 46.82%, 76.84% and 74.81%, respectively. The isolates could be classified into eight classes according to the number of drugs to which srains were resistant. The resistance spectrum of the isolates varied from 2to 9 kinds of the above drugs. The strains that were resistant to seven kinds of drugs were more than 80 percent. The resistance rates of swine origin strains to GM,AMP and KM were higher than those of poultry origin strains, while the resistance rates of poultry origin strains to TC, CMP, SM and CIP were higher remarkably than those of swine origins. The frequency of resistance increased from 2000 to 2003,which was from 67.33% to 90.58% for CMP, 71.29% to 84.81% for AMP, 73.76%to 80.10% for SM and 61.88% to 88.48% for CIP. At the same time, the resistance rate of GM decreased from 61.39% to 53.92%. Thirty-seven strains (95 %) could make all the Kunming mice die within 72 h injected intraperitoneally with the culmice three times in vivo. The pathogenicity of wild strains with drug resistance acquired naturaly to mice did not decline.

Objective To observe the application effect of point massage combined with physiotherapy in the treatment of insomnia for community elderly.Methods 30 elderly patients with insomnia who accepted point massage combined with physiotherapy were selected as observation group,and 30 elderly patients treated in tertiary hospital with insomnia who took alprazolamum for treatment were selected as control group.The SRSS score of the two groups before and after treatment,as well as treatment efficacy were compared.Results The insomnia treatment efficacy of the observation group was higher than that of the control group(Z =2.45,P <0.05).The SRSS rating of the observation group before treatment was (30.76 ±7.1 9)points and after treatment was (1 5.43 ±3.95)points.The SRSS rating of the control group before treatment was (32.33 ±6.98)points,which after treatment was (21 .93 ±6.73)points.The SRSS scores of both two groups after treatment were decreased (t =1 0.23,5.87,all P <0.05),with obvious differ-ences.The SRSS scores of the observation group decreased more obviously and was lower than that of the control group (t =4.56,P <0.05).Conclusion The point massage combined with physiotherapy can effectively improve insomnia of the elderly,which is also with advantages of easy operation and no obvious side effects and high cooperation degree etc,and it is suitable to be promoted in community.

Objective To investigate the degree of pain for patients with esophageal cancer undergoing surgery, evaluate the influence of pain on life quality, and provide evidence for therapy and nursing. Methods The SF-MPQ and BPI were used to measure the quality and intensity of the pain, and the influence on the life quality. Results Non-manual workers feeled more pain than the manual workers; the degree of the patients' education background positively correlated with PRI emotional score;postoperative pain did not correlated with ages and gender. The more pain, the worse influence on life quality. The pain affected the daily life, emotions, locomotor ability, sleep and enjoyment of life significantly. Conclusions There still existed serious pain in patients with esophageal cancer underwent surgery,and the pain affects the quality of life and postoperative treatment. Nurses should pay close attention to pain management, adopt more accurate evaluation so as to guide the pain control and improve the life quality.

The effects of pioglitazone on the expressions of hypoxia-inducible factor-1 α (HIF-1 α) and vascular endothelial growth factor (VEGF) in renal tissues of diabetic rats were observed. Diabetic rat model was established by feeding high-carbonhydrate-fat diet and injecting streptozotocin. After the treatment with pioglitazone, the kidney index, 24 h urinary albumin, blood urea nitrogen, serum creatinine, fasting blood glucose, fasting insulin, homeostasis model assessment for insulin resistance (HOMA-IR), total cholesterol,triglycerides, and low-density lipoprotein-cholesterol of diabetic rats were significantly lower than those of untreated ones, high-density lipoprotein-cholesterol was increased, the expressions of HIF-1α and VEGF in renal tissue were decreased ( all P＜0. 01 ). It suggested that pioglitazone may improve renal function and the balance of glucose-lipid metabolism in diabetic rats via down-regulating HIF-1/VEGF pathway.

BACKGROUND:There are myoblasts in human embryonic skeletal muscle. It remains poorly understand whether myoblasts in vitro can form myotube and what are the corresponding markers for identifying myoblasts and myotubes. OBJECTIVE:To investigate whether in vitro cultured myoblasts from human embryonic skeletal muscle can form myotube and whether they can express neural markers. METHODS:Human embryonic muscle-derived myoblasts were cultured in serum-containing medium. When the primary culture was established, cultured cel s were identified with immunocytochemistry for neural markers, such asβ-tubulin markers (desmin, myogenin, smooth muscle actin and myosin). RESULTS AND CONCLUSION:A population of myoblasts could migrate from human embryonic muscle tissues. They could express the markers for skeletal muscle such as desmin and myogenin, and they could express neuron specific enolase, nestin and neurofilament 200. They could form myotubes in vitro, and myotubes expressedβⅢ-tubulin, neurofilament 200 and glial fibril ary acidic protein. The data support the hypothesis that myoblasts from human embryonic muscle express neural markers and muscle markers, and cultured myoblasts and myotubes expressed neuron specific enolase,β-tubulin Ⅲ, nestin, neurofilament 200 and glial fibrillary acidic protein. This indicates that these markers could not be used for cel identification of trans-differentiation study from muscle origin to nervous system.

Objective To explore the effect and its possible mechanism of human umbilical cord mesenchymal stem cells (hUCMSCs) on bacterial growth in bronchoalveolar lavage fluid (BALF) of newborns with ventilator associated pneumonia (VAP).Methods Newborns admitted to neonatal intensive care unit (NICU) in Guangdong Women and Children Hospital Affiliated to Guangzhou Medical University from June 1,2012 to December 31,2012 were reviewed.The inclusion criteria were:(1)Positive BALF culture results.(2) Requirement of mechanical ventilation with tracheal intubation.(3) Diagnosed as ventilator-associated pneumonia.Two pieces of BALF samples of newborn were collected and randornaly divided into experimental and control group.hUCMSCs were added into the experimental group,while the same volume of conditioned medium was added into the control group.Both groups were incubated for six hours in humidified CO2 incubator at 37 ℃,then,bacterial growth was assessed by colony forming unit (CFU) counts.Levels of the antimicrobial peptides (Cathelicidin/LL-37 and human HBD-2) were determined by enzyme-linked immunosorbent assay and Western blot.Paired t-test was used for statistical analysis.Results Among the culture results of 31 newborns,there were Klebsiella pneumoniae (6 cases,19.3％),Stenotrophomonas narrow food aeromonas (6 cases,19.3％),Hemolytic staphylococci (5 cases,16.1％),Escherichia coli (3 cases,9.7％),Bacterial meningitis septicemia Elizabeth Platinum (3 cases,9.7％),Acinetobacter baumannii (3 cases,9.7％),Pseudomonas putida (2 cases,6.4％),Pseudomonas aeruginosa (1 case,3.2％),Staphylococcus aureus (1 case,3.2％) and Enterobacter cloacae (1 ease,3.2％).The CFU counts in experimental group were much less than those in control group [(2.60±0.67) ×104] CFU/ml vs [(1.18±0.32) ×105] CFU/ml,(t=-20.19,P＜0.01).Levels of Cathelicidin/LL-37 and HBD-2 in experimental group were higher than those in control group [Cathelicidin/LL-37:(8.98 ± 3.22) ng/ml vs (3.18 ± 1.57) ng/ml,t =17.79,P ＜ 0.01 ;HBD-2:(379.87±11.74) pg/ml vs (39.89±2.86) pg/ml,t=37.62,P＜0.01].Conclusions hUCMSCs had antimicrobial effect on bacterial growth in BALFs from ventilator associated pneumonia possibly by the secretion of antimicrobial peptides (Cathelicidin/LL-37 and HBD-2).

BACKGROUND:To decrease operation amount of finite element analysis and increase its clinical practice,previous studies explored the material properties and 10 kinds of material attributes were assigned,which met the requirements of finite element analysis.Moreover,it can be used to calculate bone density.OBJECTIVE:To explore a method for measurement of bone density based on three-dimensional reconstruction and finite element analysis.METHODS:A total of 11 specimens of femoral superior segment were selected.The mass of control group was firstly measured.The experimental groups were treated with thin-slice high resolution CT scan and three-dimensional reconstruction in Mimics 10.0,volume meshing in Ansys,assigned with 10,100 and 400 kinds of material attributes Mimics,exported to Ansys to calculat the volumes of the block elements of every types of material attributes.The mass and the density of the specimens was harvested according to the empirical formula concerning the gray value and the bone density.All results were treated with one-way ANOVA.RESULTS AND CONCLUSION:One-way ANOVA showed that there were no significant differences between control group and experimental groups assigned with 10,100 and 400 kinds of material attributes (P＞0.28),and there were no significantly among the experimental groups (P＞0.8).Results show that the method was able to measure the mass and the density of bone quantitatively,as well as the proportion between compact bone and cancellous bone;to assign 10 kinds of material attributes to three-dimensional model of femur could match the needs for measurements.The results can be used as an initial preparation for the unification of bone density and finite element analysis for osteoporosis.

BACKGROUND:Based on previously theoretical derivation,it thought that assignment with 10 kinds of material attributes to three-dimensional model of bone can match the needs of finite element analysis,however,whether the results is consistent with actual needs to be validated by experimental results.OBJECTIVE:Twelve specimens of femoral superior segment were used for finite element analysis,which were verified with results of biomechanical testing,to explore a reasonable method for material assignment of bone.METHODS:All 12 specimens of femoral superior segment were treated with CT scan,three-dimensional reconstruction in Mimics 10.0 and volume meshing in Ansys.The finite element analysis group was divided into 2 kinds (compact bone and cancellated bone),10,50,100,200,400 kinds of material attributes groups based on the gray value.All models were assigned with material attributes and tested in Ansys for mechanics data of nodes on surface of femoral neck.In biomechanical testing group,12 specimens of femoral superior segment were treated with compressed testing to harvest mechanics data of measuring point same as that of finite element analysis group.RESULTS AND CONCLUSION:The one-way analysis of variance showed that the differences between the biomechanical testing group and finite element analysis group of 2 kinds of material attributes had no obvious significance (P=0.082).Compared to the 10,50,100,200,400 kinds of material attributes group,the difference had no significance (P > 0.39).However,the differences between the 2 kinds of material attributes and the 10,50,100,200,400 kinds of material attributes in the finite element analysis group were obviously difference (P< 0.05),which was no difference in the 10,50,100,200,400 kinds of material attributes (P>0.9).The results demonstrated that to assign 10 kinds of material attributes to three-dimensional model of bone can match the needs of finite element analysis.