ObjectiveTo study the effect of heat stress on the cytoskeleton and cell cycle of human umbilical vein endothelial cell (HUVEC) in vitro.Methods HUVEC was cultured in vitro in 5%CO2 medium at 37℃ (control group) or 43℃ (heat stress group) for 1 hour. Coomassie brilliant blue R-250 staining was used to determine the effect of heat stress on the cytoskeleton. The cells in heat stress group were subsequently cultured at 37℃in 5%CO2 medium after heat stress for 1 hour, and cell cycle of HUVEC was determined at 0, 6, 12, 18 and 24 hours with flow cytometry.Results Under light microscopy normal cytoskeleton was observed in control group, but thicker and shorter cytoskeleton was found after a rise of temperature, and stress fibers were found in heat stress group. The DNA content of HUVEC at all time points in G0/G1 stage was 38.07%-55.19% after heat stress. The DNA content in control group was 48.57%, and it was 54.06%, 55.19%, 48.23%, 38.07%, and 41.03% at 0, 6, 12, 18, 24 hours in G0/G1 stage in heat stress group. DNA content in S phase was 35.33%-48.18%. The DNA content in control group was 44.62%, and it was 35.33%, 39.50%, 42.50%, 48.18%, and 47.99% at 0, 6, 12, 18, 24 hours in S stage in heat stress group. DNA content in G2/M phase was 5.31%-13.75%. The DNA content in control group was 6.81, and it was 10.61%, 5.31%, 9.27%,13.75%, and 10.98% at 0, 6, 12, 18, 24 hours in G2/M stage in heat stress group. It was demonstrated that compared with control group, the DNA content in G0/G1 stage was significantly increased when the HUVEC were separated from heat stress within 6 hours, and it recovered at a similar level as control group at 12 hours.Conclusion Heat stress can change the cytoskeleton of HUVEC, and cause stagnation at G0/G1 stage in cell cycle.

BACKGROUND:Fibrin glue introduced into calcium phosphate cement has not been confirmed whether this way could overcome the compressive limits and the low degradation of calcium phosphate cement and to modify the biological properties of calcium phosphate cement. OBJECTIVE:To explore the mechanical and biological properties of calcium phosphate cement/fibrin glue at different powder/liquid ratio for bone regeneration in vitro. METHODS:Calcium phosphate cement and fibrin glue were mixed at ratios of 1:1, 3:1, 5:1 (mL/g), and the pure calcium phosphate cement served as controls. Setting time, scanning electron microscope and the biomechanical test were used to analyze the composite scaffold structure, physical performance and the mechanical properties. Passage 3 osteoblasts were respectively inoculated on the material surface of the four groups, and pure cells served as blank controls. celladhesion, proliferation and alkaline phosphatase activity were observed. RESULTS AND CONCLUSION:The initial and final setting time of calcium phosphate cement/fibrin glue at 1:1 and 3:1 (mL/g) was higher than that in the control group (P<0.05), while the initial and final setting time of calcium phosphate cement/fibrin glue at 5:1 (mL/g) was lower than that of the control group (P<0.05). Scanning electron microscope showed smoother and denser surface of composite scaffolds compared with the pure calcium phosphate cement. The aperture of the composite scaffolds was decreased with the increasing concentration of fibrin glue. The compressive strength of composite scaffolds at 3:1 and 5:1 was higher than that of the control group (P<0.05), while the modulus of the composite scaffolds at 1:1, 3:1, 5:1 was higher than that of the control group (P<0.05). celladhesion, proliferation and alkaline phosphatase activity showed no difference among the three composite scaffold and control groups, but al higher than the blank control group (P<0.05). These findings indicate that fibrin glue introduced into calcium phosphate cement can overcome the low-strength limits of calcium phosphate cement, and maintain the good biological properties of calcium phosphate cement for bone regeneration.

Objective To study the effect of heat stress on the permeability,cytoskeleton and cell cycle of human skeletal muscle cell (HSKMC).Methods The HSKMC membrane permeability was detected by calcium ion inflow with flow cytometer,the cytoskeleton was stained by CBB 250,and the cell cycle was determined by flow cytometer.Results After 1 h of heat stress on human HSKMC cells under different temperature gradient,the median level of calcium ion was 91.63 in 43 ℃ heat stress group compared with 22.98 in 37 ℃ control group.As temperature increased,thicker and shorter cytoskeleton and stress fiber were shown under the high power lens of microscope.The DNA expression of skeleton cells at G0/G1 stage was 44.13-62.98 in groups under heat stress.Compared with normal control group,DNA expression was much higher in heat stress group,when HSKMC was cultured under 37 ℃ temperature for another 18 h,it kept decreasing DNA expression to a similar level as control group.Conclusions Heat stress can cause calcium iron inflow resulting in intracellular calcium overload,and affect the cytoskeleton leading to loss of normal web ordered arrangement and increased gap in HSKMC cells,which give rise to blocking cell cycle into G0/G1 stage.

Objective To discover the best physical training way including frequency and intensity each day ,and to guide pa‐tients to get the best cure .Methods Totally 60 chicken ,which were randomly divided into 3 groups ,20 for each ,the deep flexor tendon of the third left toes were found ,repairing after cut them off .Flexor tendon proximal were found by making knees incision then indwelling long silk line .External fixation bandage were used .Their left toes were accepted physical training ,Training stand‐ards :6 pulling backs each time ,toe flexion 1/4 arc ,but less than 2 N .For group A ,exercised once a day ,twice a day for group B , three times a day for group C .Right ones were consider as control group weren′t processed after operation .28 days later ,tendon distance was recorded by using 2 N pulling forces at tendon proximal with aspiring balance .Gross specimen was observed and histo‐logical specimens using Tang Jinen classification method to classify adhesions ,and the dates were analyzed by statistics SPSS 19 .0 . Results Group A remaining of 18 ,rate of 5 .56% ,grade Ⅰ of 1 ,Ⅱ of 2 ,Ⅲ of 4 ;Ⅳ of 10 .Group B remaining of 17 ,rate of 5 .88% , grade Ⅰ of 12 ,Ⅱ of 2 ,Ⅲ of 1 ,Ⅳ of 1 .group C remaining of 16 ,15 toes broke again ,fracture rate was 93 .75 % ,grade Ⅰ of 1 ,con‐trol group were grade Ⅳ .Between group A and B there were no significant differences in degree of adhesion(P> 0 .05) .Group A and Cs′ differences were statistically tested (P< 0 .05) .So were group B and C (P< 0 .05) .The average sliding distance for group A was (3 .01 ± 1 .58) mm ,(6 .72 ± 2 .02) mm for group B ,group C only got one sample ,8 .60mm ,and it was out of statistics .Be‐tween group A and B ,difference resulting from its tendon sliding after statistical tests(P< 0 .05) .For fracture rate ,Group A and B no significant difference was found(α′ = 0 .012 5 ,PAB > α′) .Conclusion Physical training can reduce chicken′s tendon adhesion effectively .

Objective To reverse P-glycoprotein-mediated muhiple drug resistance using RNA interference(RNAi)in cultured rat astrocytes.Methods Astroeytes overexpressing P-glyeoprotein induced by coriaria lactone were transfected with the short-hairpin RNA expression vector-p-SIREN shuttle designed to target Mdrl mRNA.The mRNA level of Mdrl gene was evaluated by real-time PCR;the P-glycoprotein was examined by immunocytochemistry and image analysis,meanwhile the rhodamine efflux was assessed by flow cytometry.Results The astrocyte model overexpressing P-glycoprotein were established and successfully transfected with the short-hairpin RNA expression vector-p-SIREN shuttle.The mRNA level of Mdrl gene was knocked down by 67.70%(P

BACKGROUND: The nutritional support, as well as the water and electrolyte balance during the perioperative period in the renal transplantation recipients at diuresis stage are important to the functional restoration of transplanted kidneys.OBJECTIVE: To explore the method and opportunity of the nutritional support and the handling of the water and electrolyte balance in perioperative period of renal transplantation.DESIGN, TIME AND SETTING: A retrospective clinical analysis was performed in the Department of Urology, Xijing Hospital from June 2003 to June 2007.PARTICIPANTS: Ninety-six patients of chronic renal failure underwent allograft renal transplantation. They comprised 59 males and 37 females, aged 17-67 years, with a mean of 35.7 years.METHODS: The perioperative physiological features of the renal transplantation recipients were summarized retrospectively. The recipients' condition during the perioperative period was divided into two stages at the opening point of allograft blood current. The vital signs of the patients maintained at a stable level before operation. All patients received blood transfusion since the operation began, and were supplemented with albumin before opening the vessels. Urinary production exceeding 100 mL per hour indicated the beginning of fluid replacement, which was a simplified transfusion for the patients at diuresis stage following renal transplantation.MAIN OUTCOME MEASURES: Blood inosine, urea nitrogen, electrolyte, blood sugar and urine of the patients were detected at one day postoperatively.RESULTS: During 12-16 hours postoperatively, the urinary production was 260-1 200 mL, average 520 mL per hour. Blood routine test showed 8 cases developed mild hyponatremia, accounting for 8.3%, 3 cases occurred high potassium and healed after renal functional recovery, 1 case presented low potassium and healed with supplement therapy. There were no abnormal changes of blood chlorine. The blood glucose among 21 cases (21.9%) was higher than the normal level, and recovered following hormone maneuver. The electrolytes and blood glucose were detected to be normal in other patients, without any case with low calcium or magnesium. The urine specific gravity arranged during 1.010-1.015.CONCLUSION: The colloid such as erythrocytes, blood plasma and albumin should be mainly infused before the opening of allograft blood current. And the water and electrolytes is recommended to administrate promptly and regularly during the diuresis stage. The healing of the stoma benefits from the adequate nutritional support. The metabolic acidosis still should be prevented when the urinary production returns normal.

BACKGROUND:The laparoscopy is superior to open surgery for being less invasive, inducing mild stress reaction and allowing quick recovery after operation, however the effects of laparoscopy on perioperative serum cytokine levels are controversial, and only a few studies discuss these effects among pediatric patients.OBJECTIVE: To compare the changes in perioperative cytokine levels and their clinical significance in pediatric patients undergoing laparoscopy and open surgery.DESrGN: Non-randomized concurrent controlled observation.SETTTNG: Department of Urology in Xijing Hospital of the Fourth Military Medical University of Chinese PLA.PARTICT PANTS: From May 2004 to December 2006, 135 pediatric patients for elective operation were recruited from Department of Urology in Xijing Hospital of the Fourth Military Medical University of Chinese PLA. Sixty-five patients were scheduled for laparoscopic surgery while the remaining 70 patients for open surgery.METHODS: Serum levels of interleukin-1 beta (IL-1β), IL-6, IL-10 and tumor necrosis factor-alpha (TNF-α) were measured at 24 hours before operation, and 3, 24, 48 hours after operation respectively. Duration of hospitalization time of all the children was also recorded.MAIN OUTCOME MEASURES: Levels of IL-1β, IL-6, IL-10 and TNF-α of all the patients were measured 24 hours preoperatively, and 3, 24, 48 hours postoperatively.RESULTS: All the 135 cases were included for statistical analysis. ①There were no significant perioperative changes in cytokine levels after laparoscopic surgery (P ＞ 0.05). In the open surgery group, IL-1β and IL-6 levels increased significantly at 3 and 24 hours after operation (P ＜ 0.05), and normalized within 48 hours postoperatively. No significant perioperative differences were found in IL-10 and TNF-α levels (P ＞ 0.05). The levels of IL-1β and IL-6 were significantly higher in the open surgery group than in the laparoscopic surgery group (P ＜ 0.05). ②Duration of hospitalization was shorter in the laparoscopic surgery group than in the open surgery group [(3.5±1.0), (7.5+1.5) days, P＜ 0.05].CONCLUSTON: Pediatric patients undergoing laparoscopic surgery had less perioperative changes in cytokine levels and quicker recovery.

Objective To establish a method of measuring adipose content and fat distribution of the thigh in normal glucose tolerance (NGT) subjects,and to investigate its relation to insulin resistance.Methods Insulin sensitivity was evaluated by hyperinsulinemic and euglycemic clamp technique,and femoral adipose content and fat distribution were determined by MRI in 30 individuals with NGT including 15 with normal weight and 15 overweighted or obese subjects.Results Compared to normal weight group,the subscutaneous adipose tissue of thigh (SCAT) [(176.7?21.6) cm~2 vs (115.0?12.8 ) cm~2,P＜0.05],adipose tissue of thigh beneath the fascia (SFAT) [(75.4?4.4 ) cm~2 vs (57.5?4.7 ) cm~2,P＜0.01] and intermuscular adipose tissue (IMAT) [(28.3?3.2) cm~2 vs (14.5?1.1 ) cm~2,P＜0.01] were greater in overweight/obesity group.Overweight/ obesity group had lower insulin sensitivity( glucose disposal rate under steady state of hyperinsulinemic euglycemic clamp:4.54?0.43 vs 7.88?0.75,P＜0.01).SFAT and IMAT were significantly correlated with insulin sensitivity.SFAT showed the most marked correlation with insulin sensitivity.Multiple stepwise regression analysis showed that the increased SFAT played a pivotal role in insulin resistance.Conclusion The adipose content and fat distribution are highly correlated with insulin sensitivity and the adipose tissue of thigh beneath fascia may play the most significant role in insulin sensitivity.

BACKGROUND:There is a general consensus that patients undergoing joint arthroplasty surgery wil be in hypercoagulable state and easily to induce deep vein thrombosis. Thromboelastography is a new kind of method to monitor blood coagulation state, but not widely used in orthopaedics. No final conclusion has yet been reached on whether we can guide the clinical prevention of deep vein thrombosis and medication through using thromboelastography to monitor perioperative coagulation state of patients treated with joint arthroplasty. OBJECTIVE:To investigate the correlation between thromboelastography and routine coagulation functional tests, and evaluate the clinical application value of thromboelastography in monitoring the perioperative coagulation state of patients treated with joint arthroplasty.
METHODS:A total of 204 patients who treated with joint arthroplasty at First Affiliated Hospital of Soochow University from November 2014 to August 2015 were retrospectively analyzed. The thromboelastography, routine coagulation, platelet and other data before and after the replacement were respectively col ected. The correlative analysis was conducted between the thromboelastography result and the results of conventional coagulation test, that is, routine coagulation and platelet count. RESULTS AND CONCLUSION:In total knee arthroplasty group, activated partial thromboplastin time and reaction time showed good consistency (φ=0.713, Kappa value=0.647);Prothrombin time had moderate correlation and general consistency with reaction time (φ=0.392, Kappa value=0.362);Coagulation time and fibrinogen had moderate correlation and consistency (φ=0.392, Kappa value=0.488);Aggregates formation rate (αangle) and fibrinogen had moderate correlation and consistency;the remaining parameters had poor correlation and consistency. In total hip arthroplasty group, there was a weak correlation and consistency between the reaction time, activated partial thromboplastin time and prothrombin time;the other correlations were poor. However, there was a higher proportion of consistent clotting trend between some parameters of thrombelastography and routine coagulation. In total hip arthroplasty group, the consistent proportion of coagulation time and fibrinogen accounted for 67.6%;the consistent proportion of aggregates formation rate (αangle) and fibrinogen accounted for 78.3%. These results suggest that thromboelastography and routine coagulation tests have some correlations and consistency. Thromboelastography parameters have more consistent tendency on the data. Thrombelastography can serve as an auxiliary mean to monitor coagulation state of perioperative joint arthroplasty.

OBJECTIVETo assess the influence of polymorphisms of methylenetetrahydrofolate reductase (MTHFR) gene on response to antidepressant treatment.

METHODSTwo hundred and eight one Chinese Han patients have received single antidepressant drugs for at least 6 weeks, among whom 275 were followed up for 8 weeks. Hamilton depression scale 17 (HAMD-17) was used to evaluate the severity of depressive symptoms and therapeutic effects. Single nucleotide polymorphisms (SNPs) of the MTHFR gene were determined using gene chips. Associations of single loci and haplotypes with response to treatment were analyzed using an Unphased 3.0.13 software.

RESULTSNo significant differences in gender, age, year of education, family history, episode times, and antidepressant agents were found between responders and non-responders (all P U+003E 0.05), while the baseline scores of HAMD-17 was significantly different(t=2.891, P=0.004). There was also no significant difference between age, years of education, family history, baseline scores of HAMD-17 and antidepressant agents between remitters and non-remitters (both P U+003E 0.05), while proportion of male patients was significantly higher in non-remission group than remission group (t=2.381, P=0.018), and episode times in non-remission group was significantly higher (t=-1.983, P=0.049). Single locus association analysis has found no significant association between SNPs rs1801131 and rs1801133 in the MTHFR gene with antidepressant response (P U+003E 0.05). On the other hand, haplotype A-C of MTHFR gene (rs1801131 and rs1801133) was significantly associated with antidepressant response in total group (U+03C7 2=11.39, P=0.0007), male subgroup (U+03C7 2=8.767, P=0.003) and serotonin noradrenaline reuptake inhibitors (SNRIs) subgroup (U+03C7 2=10.51, P=0.001).

CONCLUSIONParticular haplotype of MTHFR gene may be related with antidepressant effect, in which the haplotype (rs1801131, rs1801133) A-C type may be associated with better antidepressant efficacy, particularly in males and patients receiving SNRIs drugs.

Adult ; Alleles ; Antidepressive Agents ; therapeutic use ; Depressive Disorder, Major ; drug therapy ; genetics ; Female ; Haplotypes ; Humans ; Male ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Middle Aged ; Polymorphism, Single Nucleotide ; Serotonin Uptake Inhibitors ; therapeutic use ; Sex Factors ; Treatment Outcome ; Young Adult