Objective To evaluate free or pedicled perforator flaps for repairing chronic osteemylitis with soft-tissue defect in the distal lower extremity. Methods From May of 2006 to October of 2007, 28 consecutive patients of chronic osteomylitis with soft-tissue defect in the distal lower extremity underwent surgical debridement and reconstruction with free or pedicled perforator flaps. There were 13 free flaps. The free anterolateral thigh flaps were used in 2 cases to repair the soft defects in the front of leg, 3 cases in the front of the malleolus, 2 cases in the dorsum of foot, 2 cases in the heel. The free lateral crural flaps nourished by perone al artery were used in 4 cases to repair the soft defects in the dorsum of foot. There were 15 pedicled flaps. Posterior tibial artery perforator flaps were used in 4 cases to repair the soft defects in the front of leg, and 2 cases in the medial malleolus. Lateral retromalleolar perforator flaps nourished by peroneal artery were used in 6 cases to repair the soft-defects in the heel, 1 case in the lateral malleolus and 1 case in the dorsum of foot, the first dorsal metatarsal artery perforator flap was used to repair the proximal dorsum of hallux. The wound was closed with irrigation-suction in 7 cases and with vancomycin-impregnated gelatin in 8 cases. Results All 27 flaps were successfully survived except insuffcient vein refluence in 1 posterior tibial artery perforator flap, which resulted in a superficial necrosis and healed spontaneously. The follow-up period from 6 months to 2 years revealed that recurrence developed in two diffuse type patients and both were treated once and twice with success, respectively. The others healed without any signs of recurrences. No debulking procedure was necessary in any case. Secondary bone graft was performed in 3 cases. All patients were ambulatory and fully weight-bearing with normal clinical parameters at the time of last review. According to the evaluating criteria for the treatment of foot disease, the mean score was 84.5. Conclusion Free or pedicled perforator flap has been shown to be well vascularised, and it is feasible for the treatment of chronic osteomyelitis with soft-tissue defect in the distal lower extremity.

BACKGROUND:Preliminary experiments have demonstrated that rat liver homogenate supernatants can induce the morphological changes of human umbilical cord mesenchymal stem cells. However, little is known about whether the induced cells have some phenotypic and functional features of hepatocytes.
OBJECTIVE:To investigate whether human umbilical cord mesenchymal stem cells have some phenotypic and functional characteristic of hepatocytes after being induced by liver homogenate supernatants.
METHODS:Passage 3 human umbilical cord mesenchymal stem cells were used and divided into control group (cells were cultured in basic culture medium) and liver homogenate supernatant group (cells were cultured in liver homogenate supernatants for 3, 5, 7 days). Meanwhile, positive control group (QSG-7701 human liver celllines) and negative control group (simple liver homogenate supernatants) were set up. The protein and mRNA level of hepatocyte markers, alpha-fetoprotein, cytokeratin 18 and tryptophan 2,3-dioxygenase enzyme, were detected at different time points.
RESULTS AND CONCLUSION:After inducement, the stem cells of fusiform shape began to lose their sharp edges and progressively shrunk, and then they changed into hepatocyte-like cells with the morphous of triangle, polygon and anomalism shape. Compared with the control group, the protein and mRNA level of alpha-fetoprotein, cytokeratin 18 and tryptophan 2,3-dioxygenase enzyme significantly increased time dependently after inducement with liver homogenate supernatants (P<0.01). This study demonstrated that human umbilical cord mesenchymal stem cells are able to differentiate into hepatocyte-like cells in vitro that possess some functions of liver cells.

Objective To investigate the role of bacterial flagellin and CD98 in ulcerative colitis (UC).Methods A total of 60 first episode patients with active UC were recruited,including 30 mild and 30 moderate to severe UC cases.The serum of 30 healthy volunteers and normal intestinal tissues surgically removed from 15 colon cancer patients (more than 5 cm away from surgical margins) were collected as control.The content of bacterial flagellin antibodies in peripheral blood were measured with enzyme-linked immunosorbent assay (ELISA).The expression of CD98 in peripheral blood T lymphocyte was measured by flow cytometry (FACS).The expression of bacterial flagellin protein in intestinal mucosa and CD98 in intestinal epithelial basement membrane was tested by immunohistochemistry (IHC).The comparison between two groups was performed with the SNK-q method,the R×C table x2 test was used to analyze the counted data,and the Spearman correlation was used to analyze the rank materials.Results The peripheral blood concentration of bacteria flagella protein antibody of control group,mild UC group and moderate to severe group showed an upward trend,which was (7.603±2.118) pg/ml,(13.702±3.131) pg/ml and (20.813±3.004) pg/ml respectively,and the differences among groups were statistically significant (F=13.57,P＜0.01).The expression percentage of bacteria flagella protein in intestinal mucosa of the three groups also showed an upward trend,which was 3/15,56.67％ and 73.33％ respectively,and the differences among groups were statistically significant (x2 =11.553,P=0.003).The positive rate of CD98 expression in peripheral blood T lymphocytes of the three groups showed an upward trend,which was (28.42±4.31)％,(32.45±6.71)％ and (43.40±5.09) ％ respectively,and the differences among groups were statistically significant (x2 =10.110,P=0.007).The positive rate of CD98 expression in intestinal epithelial cells of the three groups also showed an upward trend,which was 1/15,36.67 ％ and 66.67％ respectively,and the differences among groups were statistically significant (x2 =5.400,P＜0.05).There was positive correlation between the peripheral blood concentration of bacteria flagella protein antibody and the expression of CD98 in peripheral blood T lymphocytes (r=0.548,P＜0.05).Conclusion Bacterial flagellin and CD98 may be important factors causing inflammatory reaction activity in UC.

Objective To investigate the application and clinical outcomes of anterolateral thigh perforator flap bridging different superficial circumflex iliac artery flaps for the treatment of complex limb wound.Methods Data of 10 patients,who had undergone free tissue transfer of anterolateral thigh perforator flap bridging different superficial circumflex iliac artery flaps (iliac osteocutaneous flap,iliac crest flap and ilioinguinal flap) for the complex limb wound from March 2009 to January 2011,were retrospectively analyzed.There were 9 males and 1 female,aged from 21 to 57 years (average,39.7 years).Iliac osteocutaneous flap was used in 7 patients,iliac crest flap in 2 patients and ilioinguinal flap in 1 patient.The free tissue transfer was performed for hand reconstruction in 3 cases,foot reconstruction in 4 cases and leg reconstruction in 3 cases.The mean size of soft tissue defect was 20 cm × 9.7 cm.Results The mean size of the anterolateral thigh flap and the ilioinguinal flap was 17.8 cm×9.4 cm and 8.4 cm×4.5 cm,respectively.The mean volume of iliac crest was 5.4 cm×2.1 cm×0.8 cm,and the mean length of flow-through conduit was 10.5 cm.Except for the distal necrosis of iliac osteocutaneous flap in 1 case,which were treated by dressing change and skin grafting,the other flaps survived without complications.All patients were followed up for 6 to 36 months (average,12 months).The average bone union time was 3 months in the hand group,4 months in the foot group,and 4.5 months in the leg group.The disabilities of the arm,shoulder,and hand questionnaire score averaged 43,the mean Japanese Orthopaedic Association's foot rating scale was 71.3,and the mean Puno's functional score was 91.Donor site scars were mostly flat,pale and soft but widened in the thigh in 6 patients and in the ilioinguinal region in 3 patients,and numbness was reported in 2 patients.Conclusion In clinical situation that requires for coverage of complex and extensive limb defects,the free tissue transfer of anterolateral thigh perforator flap bridging superficial circumflex iliac artery flap is a better method,which can meet specific reconstructive demands with minimal donor site morbidity.However,a relative high risk has to be considered.

Objective To determine which parameters of reverse remodeling of left ventricular could become the effective indexes in evaluating the short-term therapeutic effect of cardiac resynchronization therapy(CRT)in heart failure(HF)patients.Methods CRT was performed in 26 HF patients with dysfunctions of wall motion.Serial echocardiography was practiced at baseline,one and three months after CRT.The parameters including left atrial end-diastolic diameter(LADD),left ventrieular end-diastolic diameter(LVDD),end-diastolic volume (LVEDV),end-systolic volume(LVESV),ejection fraction(LVEF)were measured and compared before and after CRT therapy.Results At one and three months after CRT,respectively,CRT was associated with reduced LADD,LVDD,LVEDV,LVESV and improved LVEF and filling time compared with those at baseline.Moreover,all these parameters had better correlations with the activities of these patients.Conclusions LADD,LVDD,LVEDV,LVESV and LVEF could become the effective indexes in evaluating the short-term effect of CRT in HF patients.The reduced diameters and volumes of left atria and left ventricle are more sensitive parameters than the improved LVEF.

Objective To investigate the effect of 17β-estradiol on ketamine-induced long-term cognitive deficits in neonatal rats.Methods 80 SD male rats aged 7 days were randomly divided into group C,V,E,K and K+E,and 16 per group.Group C was intraperitoneally injected with same volume of saline for three consecutive days,Group V was subcutaneously injected with same volume of sesame oil for three consecutive days,Group E was subcutaneously injected with 600 μg · kg-1 17β-estradiol for three consecutive days,group K was intraperitoneally injected with 75 mg · kg-1 ketamine for three consecutive days,group K+E was intraperitoneally injected with 75 mg · kg-1 ketamine in combination with 600 μg · kg-1 17β-estradiol injected subcutaneously for three consecutive days.At 2 months of age,learning and memory abilities were tested with the Mortis water maze.After Morris water naze test,ten rats from each group were decapitated and the prefrontal cortex (PFC) was isolated to detect acetylcholine esterase(AchE) activity with ELISA assay and to measure acetylcholine(Ach) level by hydroxylammonium chloride method.Results The escape latency ((40.26±2.36) s,(30.25±2.20) s,(21.55±2.42) s) and path length((1019.35±58.13) cm,(811.16±27.58) cm,(598.34±34.74) cm) of group K were more than those of group C on the third,fourth aud fifth training days (all P＜0.05),while escape latency ((29.46±2.20) s,(24.86± 2.14) s,(17.20±1.91) s) and path length((913.90±41.89) cm,(729.42±31.36) cm,(487.64±18.61)cm) of group K+E were significantly lower than those of group K(all P＜0.05).On test day 6,rats were subjected to a probe trial,ratio of time spent in the target quadrant ((24.5±2.7) ％) and the number of crossings over previous platform locations (1.9±0.5) in group K were fewer than those of group C (all P＜0.05),while ratio of time spent iu the target quadrant((42.3±3.0) ％) and the number of crossings over previous platform locations(3.5±0.5) of group K+E were more than those of group K (all P＜0.05).The AchE activity((0.69±0.04) U · mg pro-1) in rats PFC of group K was significantly higher than that of group C ((0.52±0.06) U · mg pro-1) (P＜0.05).The AchE activity of group K +E ((0.58±0.12)U · mg pro-1) was lower than that of group K(P＜0.05).The Ach level ((2.59±0.34)mg · g-1) in rats PFC of group K was significantly lower than that of group C ((4.35±0.56) mg · g-1) (P＜0.05).The Ach level of group K+E((3.88±0.61) mg · g-1) was higher than that of group K(P＜0.05).Conclusions These results indicate that ketamine impairs learning and memory abilities as rat matures,while 17β-estradiol treatment improves these impairments by inhibiting AchE activity and increasing Ach level.

Objective To explore the clinical application and value of percutaneous transhepatic gallbladder drainage (PTGD)under CT guide in high-risk emergency.Methods In all 57 old patients with high-risk acute cholecystitis in emergency,cuff-PTGD in 39 was performed and fractional step PTGD in 18 was also used.Results PTGD was successfully in all patients.After PTGD,except for 1 patient died of severe cardiac insufficiency,the abdominal pain and fever were alleviated during 72 hours,and the complications was not demonstrated.Conclusion As a safe,noninvasive and accurate method,CT-guided PTGD may relieve symptoms quickly,reduce the mortality and improve the treatment for some old patients with high risk acute cholecystitis.

Ascoviruses, iridoviruses, asfarviruses and poxviruses are all cytoplasmic DNA viruses. The evolutionary origins of cytoplasmic DNA viruses have never been fully addressed. Morphological, genetic and molecular data were used to test if all four cytoplasmic virus families (Ascoviridae, Iridoviridae, Asfarviridae, and Poxvirirdae) evolved from nuclear replicating baculoviruses and how the four virus groups are related. Molecular phylogenetic analyses using DNA polymerase predicted that cytoplasmic DNA viruses might have evolved from nuclear replicating baculoviruses, and that poxviruses and asfarviruses share a common ancestor with iridoviruses. These three cytoplasmic viruses again shared a common ancestor with ascoviruses. Morphological and genetic data predicted the same evolutionary trend as molecular data predicted. A genome sequence comparison showed that ascoviruses have more baculovirus protein homologues than do iridoviruses, which suggested that ascoviruses have evolved from baculoviruses and iridoviruses evolved from ascoviruses. Poxviruses showed genetic and morphological similarity to other cytoplamic viruses, such as ascoviruses, suggesting it has undergone reticulate evolution via hybridization, recombination and lateral gene transfer with other viruses. Within the ascovirus family, we tested if molecular phylogenetic analyses agree with biological inference; that is, ascovirus had an evolutionary trend of increasing genome size, expanding host range and widening tissue tropism for these viruses. Both molecular and biological data predicted this evolutionary trend. The phylogenetic relationship among the four species of ascovirus was predicted to be that TnAV-2 and HvAV-3 shared a common ancestor with SfAV-1 and the three virus species again shared a common ancestor with DpAV-4.

OBJECTIVETo observe the in vivo migration of human umbilical cord mesenchymal stem cells (hUCMSCs) labeled with the PKH26 red fluorescent dye after transplantation into rats with liver cirrhosis.

METHODSFrozen hUCMSCs were resuscitated and labeled with PKH26. Labeling efficiency and fluorescent maintenance time of the PKH26-1abeled cells were measured. Morphology of the labeled and unlabeled (control) cells was observed by microscopy. The cell growth curve was determined using the MTT method. The PKH26-1abeled hUCMSCs were transplanted via tail vein injection into healthy (control) rats and rats with liver cirrhosis. Migration of the PKH26-1abeled hUCMSCs observed 48 h later in frozen liver sections under a fluorescence microscope.

RESULTSThe labeling ratio of PKH26 to hUCMSCs was 100%. Growth of the labeled cells was good. The cell morphology was not significantly different between the labeled and unlabeled cells; all cells were long and spindle-like. Cell proliferation was not impacted significantly by labeling.Fluorescence was maintained for at least 20 days, as detected by in vitro analysis. After transplantation into the rats, the PKH26-1abeled hUCMSCs were mainly distributed in the area surrounding the portal vein, the blood vessels, and the false lobule of the cirrhotic liver; a small amount ofhUCMSCs were present in the spleen and lung.

CONCLUSIONPKH26 is an ideal fluorescent dye to label hUCMSCs. The PKH26 labeling technique can be used to study the migration of hUCMSCs in cirrhotic liver.

Objective To evaluate the intra- and interatrial synchronization before and after pacemaker implantation in patients with sick sinus syndrome ( SSS) by tissue Doppler imaging ( TDI) . Methods Thirty-eight patients with SSS ( SSS group ) underwent pacemaker implantation . Echocardiography was performed before and after operation . Twenty-five volunteers were chosen as control group . Routine measurement of left and right atrial diameter by echocardiography . In apical four chamber view ,the sample volume of TDI was located at right atrial lateral wall at tricuspid annulus ,interatrial septum and left atrial lateral wall at mitral annulus . The interval time between the early diastolic wave ( E wave) and the late diastolic wave (A wave) (E-AR ,E-AI ,E-AL ) ,and the duration of A wave (AR ,AI ,AL ) were measured . At the same time ,the electromechanical coupling time was measured ,namely the initiation of the P wave of the synchronous electrocardiogram to the starting point of A wave ( P-A0R ,P-A0I ,P-A0L ) , and the starting point of the P wave to the peak point of the A wave ( P-AR ,P-AI ,P-AL ) . And the right atrial electromechanical delay time ,P-A0I and P-A0R value ( T0IR ) ,P-AI value and P-AR value ( TIR ) were recorded .Other parameters include left atrial electromechanical delay time:P-A0L and P-A0I value ( T0LI ) , and P-AL and P-AI value ( TLI ) ;interatrial electromechanical delay time:P-A0L and P-A0R value ( T0LR ) ,P-AL and P-AR ( TLR ) were recorded . Results Compared with the control group ,left atrium and right atrium in SSS group enlarged before operation , E-AR ,E-AI ,E-AL ,P-A0R ,P-A0I ,P-AR ,P-AL prolonged ,while T0LR and T0LI were shortened ( all P < 0 .05 ) . Postoperative changes in SSS group patients included shortened E-AR ,E-AI and E-AL ,and prolonged T0LI and T0LR ( all P <0 .05) . Compared with the control group ,the size of left atriums and right atriums in SSS group still increased postoperation ,and the P-A0I 、P-AR and P-AL remained longer (all P <0 .05) . The same as the control group ,the postoperative AR ,AI and AL in SSS group were decreased successively ( all P < 0 .05 ) . Conclusions In SSS patients , electromechanical asynchrony exists in different part of atrium and between left atrium and right atrium . Pacemaker therapy can improve atrial electromechanical synchronicity .