Objective To discuss the functional results of percutaneons compressive screw fixation for float injury to the pubic symphysis. Methods From March 2003 to March 2007, 48 cases of float injury to the pubic symphysis were treated with percutaneons compressive screws, including 27 males and 21 females with an average age of 29.4 years. Of them, 39 eases were complicated with injury to the pelvic posterior ring. Emergency surgery was done for 13 cases, 27 cases were operated on within 3 to 7 days after injury and 8 within 7 to 14 days. Guided pins and screws were used during dosed reduction and percutaneous pelvic fixation was done under the guidance of intraoperative fluoroscopic imaging. Float injury to the pubic symphysis was amended by percutaneous fixation after dosed manipulation. Results The average operation time for the48 patients was 55 (31 to 100) min. The intraoperative bleeding averaged 20 to 30 mL. Satisfactory reduetian and fixation was achieved in 41 cases, but 7 cases had poor reduction. All the fractures healed 3 to 6 months postoperatively without infection, nonunion or injury to vessels, nerves or organs. All the patients could turn the body freely in bed the day after operation. Those without injury to the pelvic posterior ring could walk with crutches 3 days after operation. By the Orlando evaluation system for pelvic fractures, 37 eases were rated as excellent, 7 as good, 3 as fair and I as poor. Conclusions The percutaneous compressive screw fixation may decompress the pelvic hematoma, allowing early definitive fixation without the risk of additional hemorrhage. Complications associated with open posterior pelvic surgical procedures may be avoided by using percutaneons techniques.

[ABSTRACT]AIM:ToinvestigatethepromotingroleofTranswellcontactco-culturesysteminthegrowthand differentiation of single-dissociated induced pluripotent stem cells (iPSCs).METHODS:Bovine corneal endothelial cells (CECs) at passage 1~2 (P1~2) were seeded on the underside of Transwell inserts placed into culture plates and were cultured in 37 ℃and 5%CO2 for 8 h.Accutase digestion and 40μm filter process disaggregated colony-aggregated iPSCs into single-dissociated iPSCs , and the cells were seeded on the inside of Transwell inserts with CECs in medium of mTeSR 1 for 3 d and then in low-glucose DMEM supplemented with 10% FBS for 2 weeks.The characteristics and differentiation markers were evaluated by real-time fluorescence quantitative polymerase chain reaction ( qPCR ) , immunofluorescence staining, live&dead cell staining and alkaline phosphatase (ALP) staining.The group of iPSCs cultured in conventional medium was used as control group 1.The group of single-dissociated iPSCs co-cultured with CECs was set as experimental group, while single-dissociated iPSCs without co-culture were as control group 2.RESULTS: The bovine CECs showed typical hexagonal cobblestone shape .iPSCs showed colony-like growth , while became single-dissociated cells after Tran-swell contact co-culture with bovine CECs for 3 d.The single-dissociated iPSCs positively expressed the undifferentiated markers, Nanog and Oct4.The mRNA expression levels of Nanog , Oct4 and Sox2 between experimental group and control group 1 were both positive and had no statistical significance difference (P>0.05).The dead cells in experimental group decreased significantly, and there was statistically significant difference compared to control group 2 (P<0.01).After 14 d of induced differentiation co-culture , the single-dissociated iPSCs showed rather uniform polygonal morphology , increased dimension and no obvious colony existence .Negative ALP staining, positive immunofluorescence staining for ZO-1, AQP1 and CD31, and negative for CD34 and CD133 were also observed.The results of qPCR showed that the mRNA expression of Oct4, Nanog and Sox2 significantly decreased , and had statistically significant difference compared with control group 1 (P<0.01).CONCLUSION: When co-cultured with bovine CECs, iPSCs morphologically changed to endothelial-like cells and expressed some markers of CECs .Transwell contact co-culture system not only enhances the growth of single-dis-sociated iPSCs , but also promotes their differentiation .

[ ABSTRACT] AIM:To investigate the effect of maxadilan, which specifically activates pituitary adenylate cycla-se-activating polypeptide type I receptor (PAC1 receptor), on the proliferation, apoptosis and differentiation potential of human adipose-derived stem cells ( ASCs) .METHODS:ASCs from human adipose tissue were isolated by enzymatic di-gestion and cultured.ASCs were confirmed by the analysis of the markers for cell phenotypes by flow cytometry ( FCM) and adipogenic/osteogenic induction.The effect of maxadilan on ASCs viability was analyzed by CCK-8 assay and FCM.ASCs were irradiated by ultraviolet C ( UVC) at 254 nm and the absorbance of apoptotic ASCs induced by various doses of UVC was measured by CCK-8 assay.ASCs were exposed to 702 J/m2 UVC for 24 h to induce apoptosis.The effect of maxadilan on ASC apoptosis was analyzed by FCM and the determination of caspase 3 and caspase 9 levels.RESULTS:Adipose-de-rived stem cells were confirmed by the detection of the positive expression of cell phenotypes including CD29, CD44, CD59 and CD105 by FCM.The data of CCK-8 assay revealed that ASCs treated with maxadilan (80 nmol/L) had the strongest ability of proliferation.The data of FCM also demonstrated that the addition of 80 nmol/L maxadilan to ASCs in experimen-tal group markedly improved the proliferation capacity of the cells compared with control group (P<0.05).The apoptosis of ASCs exposed to 702 J/m2UVC was dramatically inhibited by the treatment with maxadilan (80 nmol/L).Such process involved the caspase signaling pathway including caspase 3 and caspase 9.There was statistical significance (P<0.05) between experiment group ( ASCs irradiated by UVC and supplemented with maxadilan) and control group ( ASCs only irra-diated by UVC) .Meanwhile, adipogenic and osteogenic differentiation potentials were both positive in experiment group and control group.CONCLUSION:Maxadilan promotes proliferation and inhibits apoptosis of the ASCs.The differentia-tion potential of ASCs toward adipogenic and osteogenic lineages wouldn’ t be altered by maxadilan.Maxadilan would bene-fit to growth and expansion of ASCs in vitro.

Objective: To explore the in vitro inhibition effect of Zuojin pills on 6 cytochrome P450 isoenzymes ( CYP1A2, CYP2C8, CYP2C9, CYP2C19, CYP2D6 and CYP3A4) in human liver microsomes. Methods:The water extract of Zuojin pills, Cop-tidis Rhizoma and Euodiae Fructus was respectively incubated with human liver microsomes in the presence of seven probe substrates of CYP450 isoenzymes, and seven metabolites of cytochrome P450 probe substrate ( paracetamol/CYP1A2, 6α-hydroxypaclitaxel/CYP2C8, 4-hydroxydiclofenac/CYP2C9, 4-hydroxymephenytoin/CYP2C19, dextrorphan/CYP2D6, 6β-hydroxytestosterone/CYP3A4 and 1-hydroxymidazolam/CYP3A4) were simultaneously measured by LC-MS/MS, and the inhibitory effects were evaluated with IC50 value. Results:The IC50 value of Zuojin pills on CYP2D6, CYP1A2 and CYP3A4_T was 11. 6, 77. 4 and 97. 0 μg·ml-1 , respec-tively. The other IC50 values were from 334 to 690μg·ml-1 on CYP2C8, CYP2C9, CYP2C19 and CYP3A4_M isoenzymes. The IC50 value of Coptidis Rhizoma on CYP2D6, CYP1A2 and CYP3A4_T was 5. 8, 36. 8 and 59. 2 μg·ml-1 , respectively. The other IC50 values were from 163 to 476 μg·ml-1 on CYP2C8, CYP2C9, CYP2C19 and CYP3A4_M isoenzymes. The IC50 value of Euodiae Fructus on CYPs was over 107 μg·ml-1 . Conclusion:Zuojin pills shows notable inhibitory effect on CYP2D6, and weak inhibitory effects on CYP1A2 and CYP3A4_T. Coptidis Rhizoma has similar effects on CYPs and may be the main herbal medicine in the formu-la. Therefore, much attention should be paid to the combination of Zuojin pills and the drugs metabolized by human CYP2D6 in clin-ics.

Objective To study the therapeutic efficacy of the chondroplasty with radiofrequency technique under arthroscope for osteoarthritic knees in the senile patients.Methods Thirty-six patients were divided randomly into control and test groups(in each with 18 patients).The patients suffered from osteoarthritis of knees.The control group confirmed and treated by knee arthroscope,which was subjected to debridement with routine technique.The test group was treated by chondroplasty with radiofrequency technique under arthroscope.Results The patients developed no infection or other complications.Better therapeutic results were obtained after follow up of 6 to 18 months.Conclusions Chondroplasty with radiofrequency technique under arthroscope for the osteoarthritic knees has the advantages of less injury,less bleeding,less complication and quick recovery.

BACKGROUND:With the deepen understanding on the biological function of Rho/ROCK pathway, new ROCK inhibitors continue to be discovered, and ROCK inhibitors show good promoting effects on the survival, proliferation and migration of keratocytes. Research on ROCK inhibitors wil provide more donor materials or seed cels for regenerative medicine and clinical cel transplantation. OBJECTIVE:To summarize and explore the progress in the treatment and application of corneal disease using the ROCK inhibitors Y-27632 and Y-39983. METHODS:The PubMed database and CNKI database were retrieved by computer to search the relevant literature published between 2008 to 2015 using the key words of “corneal endothelial cel, corneal epithelial cel, ROCK inhibitor, Y-39983, Y-27632” in English and Chinese, respectively. Relevant articles in line with the theme were screened and analyzed. RESULTS AND CONCLUSION:Totaly 264 papers were initialy searched. At last, 45 papers were selected. Currently there are two main ROCK inhibitors: Y-27632 and Y-39983, but both of which are stil in basic research stage and clinical testing stage. Y-27632 promotes the proliferation and activity of corneal epithelial stem cel after resuscitation; Y-39983 as a novel ROCK inhibitor can be better to inhibit Rho kinases activity than Y-27632, thereby more effectively promoting the healing of the corneal endothelium. There are many studies on the application of ROCK inhibitors in corneal treatment, but not a stable method established to obtain seed cels. Each method has its own advantages and disadvantages, and how to overcome these disadvantages and to find fast and stable access to seed cels is the future direction of development.

Induced pluripotent stem cells ( iPSCs) have been first induced from mouse fibroblasts since 2006, and the research on iPSCs has made great progress in the following years .iPS cell lines were established from different so-matic cells through DNA , RNA, protein, and small molecule compounds and various methods of transduction , making the induction of iPSCs more secure and effective , and more attractive prospect of clinical application .In this review , different somatic cell reprogramming , different levels of reprogramming , different transduction pathways , and prospect of application are discussed .

Background The construction of tissue-engineered corneal endothelium and corneal endothelial cells (CECs) therapy need abundant seed cells,so how to culture a large amount of CECs with high viability and original cell properties is an urgent issue to be solved.Objective This study was to establish three-dimensional spheroid culture of CECs and explore the cellular biological characteristics.Methods Primary rabbit CECs were isolated with trypsin and subcultured.Low attachment and shaking culture was applied to form CECs spheres.Cultured cells were identified under the inverted microscope.The surface features of the cells were examined under the scanning electron microscope.The viability of the cells were assayed by acridine orange (AO) staining and CCK-8 kit.Then CECs spheres were incubated to 6-well plate for 1 week,and immunofluorescence staining was used to identify the expression of zonula occludens-1 (ZO-1) and Na+/K+-ATPase in the cells.The cell proliferation value of spheroid culture method was compared with that of regular culture method.Results CECs grew into aggregation after cultured with the hexagonal or polygonal shape and tight connection among the cells.The cells converged into single layer and slabstone-like arrangement 1 week later.Cells migrated out of the CECs sphere and formed an uneven spherical surface.The living cells showed green fluorescence for AO with the survival rate 90％.The absorbance (A450) of the cells was 1.524±0.013 and 1.265 ±0.021 in the spherical culture group and conventional culture group,respectively,showing a significant difference between them (t =-3.436,P=0.010).The positive cells of ZO-1 and Na+/K+-ATPase showed the green fluorescence for FITC on cell membrane and blue fluorescence for DAPI on cell nucleus.Conclusions Spherical culture method maintains a high viability and proliferation ability of the cells and remains phenotype of CECs,which is superior to conventional culture method.This culture method provides better seeding CECs for the establishment of tissue engineering cornea endothelial layer and CECs therapy.

BACKGROUND:Many types of mammalian cells aggregate and display three-dimensional multicellular spheroids when they are in normal physiological conditions. In order to observe and explore cellular natural states, many researchers try to use spherical cellculture in vitro, a common three-dimensional culture pattern.
OBJECTIVE:To use three different methods for spherical culture in vitro of adipose-derived stem cells and to observe their biological features.
METHODS:Adipose-derived stem cells were confirmed by the analysis of the markers for cellphenotypes as wel as adipogenic and osteogenic differentiation potential assays. Three different methods of sphere cultures were used as fol ows:(1) ultra low attachment culture;(2) hanging-drop culture and (3) Eppendorf tube culture. The sphere formation was compared among above three methods. We used Imagej to calculate mean areas of these spheres. And we used Viability/Cytotoxicity Assay Kit for Animal Live&Dead cells to detect their vitality.
RESULTS AND CONCLUSION:(1) Adipose-derived stem cells were confirmed by the analysis of the markers for cellphenotypes, CD29, CD44, CD59 were positive, as wel as adipogenic and osteogenic differentiation potential assays were positive. The conventional monolayer cultures of adipose-derived stem cells showed spindle and cloning growth within three passages. (2) Ultra low attachment culture, hanging-drop culture, Eppendorf tube culture al could elicit adipose-derived stem cells spherical growth. However, spherical size, shape and uniformity differed depending on cellnumbers, culture time and spherical culture methods. The ultra low attachment culture was comparatively difficult to control spherical shape and uniformity of adipose-derived stem cells. But hanging-drop culture and Eppendorf tube culture were able to form even cellspheres. (3) Spherical formation of adipose-derived stem cells using our three methods displayed good cellvitality.

This study was aimed to observe the antipyresis and cholagogue effect of theFructus Gardeniae-Fructus Forsythiaeherb couple in order to explore the possible pharmacodynamic mechanism. Dry yeast suspension was subcutaneously injected into the back of rat to establish the fever model. The freeze-dried powders ofF. Gardeniae,F. Forsythiae, andF. Gardeniae-F. Forsythiaeherb couple were prepared. They were dissolved in the water for intragastric administration. SD rats were randomly divided into the normal group, model group, positive control group and Chinese medicine group. The Chinese medicine group was subdivided into the group ofF. Gardeniae,F. Forsythiae, andF. Gardeniae-F. Forsythiaeherb couple. The concentration of herbal water extract was 10 mL·kg-1 (which equaled to 3 g·kg-1 of a single crude herb). The concentration of positive control was 10 mL·kg-1. Intragastric administration of equal amount of normal saline was given to the blank group and the model group. Except rats in the normal group, rats in other groups were subcutaneously injected with 10 mL·kg-1 of 15% dry yeast suspension on the back of to establish the fever model. Electronic thermometer was used to record the body temperature of rats at 0, 1, 4, 5, 6, 7 and 8 h after the injection, respectively. Meanwhile, bile was collected from 1-1, 1-2, 2-4, 4-6, 6-8, 8-12, 12-24 h, respectively. Observation was given on changes of body temperature and bile amount of rat. The results showed thatF. Gardeniae,F. Forsythiae, andF. Gardeniae- F. Forsythiaeherb couple had certain effect to reduce the body temperature of rats. The temperature-reducing effect of the combination of both herbs was better than a single herb. TheF. Gardeniae -F. Forsythiae herb couple can reduce the body temperature of fever rat (P < 0.05, orP < 0.01). It had a little effect on the body temperature of normal rat. TheF. Gardeniae - F. Forsythiaeherb couple can promote the bile secretion, which was better than the single using of F. Gardeniae (P < 0.05). It was concluded that theF. Gardeniae - F. Forsythiaeherb couple had better temperature-reducing effect than the using of a single herb; however, there was no significant difference. But it had obvious effect on the promotion of bile secretion, which indicated the strengthening of cholagogue effect.