AIM:To investigate the role of PI3K/Akt and JAK2/STAT3 pathways in the protection of sulfur dioxide (SO2) against limb ischemia/reperfusion (I/R)-induced acute lung injury (ALI) in rats.METHODS:ALI was induced by limb I/R in the SD rats.Na2 SO3 (0.54 mmol/kg, ip)/NaHSO3 (0.18 mmol/kg, ip) as SO2 donor was injec-ted at 20 min before reperfusion.The inhibitors of JAK2/STAT3 and PI3K/Akt pathways, Stattic (3 mg/kg, iv) and LY294002 (40 mg/kg, iv), respectively, were injected at 1 h before reperfusion.Peripheral blood and lung tissues were collected for determining the contents of the cytokines, the protein levels of the molecules related to the signaling pathways, apoptosis and histopathologic changes by ELISA, TUNEL and Western blot.RESULTS:Compared with control group, the content of MDA, the activity of MPO, lung coefficient, apoptotic index, cytokine expression, and the protein levels of p-Akt and p-STAT3 in I/R group all increased significantly, and administration of Na2 SO3/NaHSO3 attenuated the damage in the lung.Besides, the results of Western blot showed that the rat lung tissues expressed p-STAT3 protein and p-Akt pro-tein.After I/R, the protein levels of p-STAT3 and p-Akt were increased.After using Na2 SO3/NaHSO3 , p-Akt was in-creased, but p-STAT3 was decreased (P<0.05).CONCLUSION:Both JAK2/STAT3 and PI3K/Akt pathways are like-ly involved in the protective effect of SO2 against limb I/R-induced ALI in rats.The activation of JAK2/STAT3 signaling pathway increases I/R injury.Reversely, the activation of PI3K/Akt signaling pathway reduces I/R injury.Besides, JAK2/STAT3 and PI3K/Akt signaling pathways may have crosstalk during I/R-induced ALI and JAK2/STAT3 pathway may have an impact on the P13K/Akt pathway.

Objective To investigate the expression and clinical significance of Smac and HtrA2 in children with acute leukemia(AL).Methods Bone marrow samples were obtained from 77 children with AL (including 32 newly diagnosed children,33 complete remission children and 12 relapsed children)and the control group of 15 children without malignant blood disease.The expressions of Smac and HtrA2 protein were measured by streptavidin/peroxidase immunoperoxidase technique(SP) in all children.SPSS 13.0 software was applied to analyze the statistical data.Results Protein Smac was detected only in some samples,but HtrA2 was detected in all samples.The levels of Smac and HtrA2 protein in newly diagnosed AL children were both higher than those of the complete remission children (x2 =17.38,F =2.36,all P ＜ 0.05) and normal controls (x2 =12.89,F =5.26,all P ＜ 0.05),there was a statistical significance,but compared with those in the relapsed children,the difference had no statistical significance (x2 =1.18,F =1.57,all P ＞ 0.05).The levels of Smac and HtrA2 protein in complete remission children were both higher than those of the normal controls,and the difference had no statistical sigmficance(x2 =1.20,F =2.23,all P ＞ 0.05).In the newly diagnosed children,the levels of Smac and HtrA2 protein in children with acute lymphocytic leukemia(ALL) were higher than those of the acute myeloid leukemia(AML),but the differences had no statistical significance(x2 =0.113,t =1.024,all P ＞ 0.05).In newly diagnosed AL children,the complete remission(CR) rate of the negative expression of Smac(Smac-,90.9％) and the low expression of HtrA2(HtrA2low,84.6％) in the level of protein were higher than those of the positive expression of Smac(Smac +,47.6％) and the high expression of HtrA2 (HtrA2high,47.4％),and there was statistical significance respectively(x2 =5.772,4.596,all P ＜ 0.05).The CR rate of Smac-HtrA2low group (100％) was higher than that of Smac+ HtrA2high group(30.8％)in the children with AL,and the statistical data were of great significance(x =9.692,P ＜0.01).The protein level of Smac in newly diagnosed AL children was correlatedwith HtrA2 (r =0.979,P ＜ 0.001).Conclusions Pro-apoptotic protein Smac and HtrA2 may be involved in and af-fected each other in the pathogenesis and progression in AL,but levels of Smac and HtrA2 protein may be not correlatedwith the types of AL.In newly diagnosed AL children,the high expression of protein Smac and HtrA2 predicts poorprognosis.

Objective To explore the influence of brief ischemic on fracture healing.Methods Two hundred and forty wistar rats (weight was 200 g and half male) were made the right tibial midpiece closed fracture model.1.0 mm K-wires were used to fix the fractures.Rats were divided into 4 groups.Group A was 24 hours,group B was 48 hours,group C was 72 hours,and group D was the control group.Tourniquet was used on the root of the thigh of right lower limb.Tourniquet was inflated 10 min,then released 10 min,and repeated 3 times to make the brief ischemic model.48 rats were made brief ischemic at 24 hours after the fracture fixed.Then group A was at every 24 hours,group B was at every 48 hours,and group C was at every 72 hours made brief ischemic.Group D at every 24 hours after the fracture fixed was used tourniquet on the root of thigh of right lower limb each time 1 hour,but the tourniquet was not inflated.Group A,B,and C were made brief ischemic 5 weeks.Group D was tourniquet treated 5 weeks.Group A and D at 1 day,3 days,5 days,and 7 days after fracture fixed were detected the serum VEGF.All groups of rats were X-ray examinated and detected of serum VEGF at every two weeks after the fracture fixed.Every two weeks,every group randomly selected 12 rats to do immunohistochemical staining.We used the Image Pro Plus software to quantitative compare the IGF-2 positive cells in the fracture area.Results X-ray results were used three points method for analyzing.The scores of group A,B,and C were significantly higher than group D at 2 weeks,4 weeks,6 weeks after fracture fixed (P ＜ 0.05).The scores between group A,B,and C were no significant statistical difference at 2 weeks,4 weeks,6 weeks after fracture fixed (P ＞ 0.05).The levels of serum VEGF of group A were significantly higher than group D at 3days,5days,and 7 days after fracture fixed (P ＜ 0.05).The levels of serum VEGF of group B and C were obviously higher than group A and D at 2 weeks (P ＜ 0.05),and there was no obvious difference between group B and C (P =0.79,P =0.90,P =0.88).The same result was found between group A and D (P =0.94,P =0.551,P =0.53).There were no significant difference between four groups on the levels of serum VEGF at 8 weeks (P =0.66).Immunohistochemical results were that group B,group C VEGF positive cells were significantly higher than group A and group D at 2 weeks,4 weeks,6 weeks (P ＜0.05).There was no significant difference in group B and group C (P =0.19,P =0.10,P =0.38),or group A and group D (P =0.07,P =0.17,P =0.18).There was no significantly different in four groups at 8 weeks (P =0.08).Conclusions Brief ischemic can promote the healing of fracture and stimulate the expression of VEGF.The mechanism may be related to the signal channels of VEGF and other biological factors.This study will give new idea to clinic for promoting fracture healing.

Objective To evaluate the effect of exogenous nitric oxide (NO) on Schistosoma japonicum in mice. Methods The emulsion of nitric oxide was performed by the reversal emulsifying method and beeswax was used as an agent absorbing NO. Each mice on the day 22 post-infection with cercariae was orally administracted with exogenous NO emulsion 0.5 ml once a day for five days. The worms of Schistosoma japonicum were collected by perfusion and counted to observe the effect of exogenous NO on Schistosoma japonicum. Results The concentration of exogenous NO was 536.2 ?mol/L. Exogenous NO could reach certain therapy efficacy with the worm reduction rate of 45.0% and the egg reduction rate of 42.7%. Conclusion Exogenous NO may be considered as a novel medicament of schistosomiasis japonica.

BACKGROUND:Previous studies have found that endogenous gaseous signaling molecules such as NO, CO, H2S and SO2 play an important role in acute lung injury;there also have other gases participation, such as carbonyl sulfide.
OBJECTIVE:To investigate the effect of carbonyl sulfide for limb ischemia-reperfusion induced acute lung injury and its mechanism in rats.
METHODS:A total of 64 Sprague-Dawley rats were randomly divided into eight groups. Control group:without treatment;model group:limb ischemia for 4 hours and then reperfusion for 2 hours. Low-, moderate-and high-dose carbonyl sulfide groups were intraperitoneal y injected with 0.2, 0.5, 1.0 mL carbonyl sulfide respectively at 20 minutes before ischemia for 4 hours and reperfusion for 2 hours. Low-, moderate-and high-dose air groups were intraperitoneal y injected with 0.4, 1.0, 2.0 mL air respectively at 20 minutes before ischemia for 4 hours and reperfusion for 2 hours. 2 hours after reperfusion, the morphological changes of lung tissues and the change of lung coefficient were observed. The expressions of tumor necrosis factor-a, interleukin-1βand interleukin-6 both in lung tissue and serum were detected by enzyme linked immunosorbent assay. Cel apoptosis was measured by TUNEL assay.
RESULTS AND CONCLUSION:(1) Compared with the control group, significant damage of lung tissue was seen, and the lung coefficient increased significantly in the model group (P<0.05). The expressions of tumor necrosis factor-a, interleukin-l and interleukin-6 both in lung tissue and plasma increased (P<0.05), and apoptotic rate increased. (2) Compared with the model group, low-, moderate-and high-doses of carbonyl sulfide could mitigate the degree of lung injury, and reduce pulmonary coefficient and apoptotic rate. The low dose showed the most obvious effect. Low-and moderate-dose carbonyl sulfide could significantly decrease expressions of tumor necrosis factor-a, interleukin-1βand interleukin-6 both in lung tissue and plasma (P<0.05). (3) No significant difference in each index was visible in the low-, moderate-and high-dose air groups compared with the model group. (4) Results suggested that low dose of exogenous carbonyl sulfide through anti-inflammatory, anti-oxidant effects plays the protective role on limb ischemia/reperfusion-induced acute lung injury in rats.

Objective To analyze the characteristics and clinical value of time intensity curve (TIC) of contrast-enhanced ultrasound (CEUS) in recurrent small hepatocellular carcinoma (RSHCC) and primary small hepatocellular carrcinoma (PSHCC). Methods Sixty-five cases of RSHCC (all lesions ≤3 cm) were devided into group B1 with 42 cases of RSHCC (≤2 years ) , and group B2 with 23 cases of RSHCC ( > 2 yeras ) and group A invloved 49 cases of PSHCC (all lesions ≤3 cm). Enhancement patterns in arterial, portal and delayed phase were evaluated respectively in three groups through CEUS and analytic software Sonoliver was applied to obtain quantitative features of CEUS in the region of interest. Receiver operating characteristic (ROC) was drawn and the area under curve (AUC) was calculated. Results CEUS showed hyper-enhancement difference in arte-rial phase in group B2 (72.4%) and group A (94.8%)(P′ = 0.008) showed statistical significance, but no sig-nificance was found in enhanced iso in portal phase (P = 0.078). Hypo-enhancement in the delayed phase in group B2 (75.9%), group A (96.6%) and group B1 (95.3%) (P′ = 0.003, P′ = 0.005). TIC showed HT difference (half time of descending) in B2 group, A group and B1 group (P′ = 0.007, P′ = 0.013) indicated statistical significance but RT, TTP, MTT(P = 0.319,P = 0.104, P = 0.461) showed no difference. AUC was 0.841 (half time of descending). Conclusions Enhancement patterns of CEUS (RSHCC) are related to recur-rent time . En hancement patterns of RSHCC (> 2 years ) is not typical so CEUS should be combined with quanti-tative analysis of TIC to provide reference for its treatment and prognosis.

BACKGROUND:Some studies have shown that blocking the venous return can promote fracture healing but there is no study about the effect of repeated transient ischemia on promoting fracture healing. OBJECTIVE:To observe the promoting effect of repeated transient ischemia on early fracture healing based on micro-CT technique. METHODS: Forty-eight Wistar rats were used to make animal models of right tibial midpiece closed fractures folowed by K-wire fixation. Rats were randomly divided into four groups: 24-hour group, 48-hour group, 72-hour group and control group. Tourniquet was used on the root of the affected thigh, inflated for 10 minutes, and then released for 10 minutes. Three above-mentioned cycles were taken as an ischemic treatment. At 24 hours after fracture fixation, the ischemic treatment was first carried out, and after that, it was done every 24 hours in the 24-hour group, every 48 hours in the 48-hour group and every 72 hours in the 72-hour group. In the control group, the tourniquet with no inflation was used once on the right lower limb every 24 hours after fixation. Al the rats were kiled at 2 weeks after modeling to take out the right tibia with removal of K-wires for micro-CT observation. RESULTS AND CONCLUSION:The trabecular bone volume and total volume in the regions of interest were significantly higher in the 24-hour group than the other three groups (P < 0.05), but the tissue mineral density and calus tissue mineral density were significantly lower in the 24-hour group than the other three groups (P < 0.05).There was no significant statistical difference in the bone volume fraction among the four groups (P > 0.05). These findings indicate that within 2 weeks after fractures, the repeated transient ischemia can promote fracture healing, which may be related to the improvement in the secretion of growth factor, revascularization and osteoblast proliferation induced by repeated transient ischemia.

ObjectiveTo assess the factors inlfuencing the therapeutic effects of INSURE technology in premature in-fants with respiratory distress syndrome (NRDS).MethodsThe clinical data from 309 infants with NRDS treated by INSURE technology were retrospectively analyzed from Jan. 2000 to Dec. 2012.ResultsIn 309 infants with NRDS, 302 infants were cured and the cure rate was 97.7%. Twenty-one infants (6.8%) needed the reintubation for mechanical ventilation within 72 h. The difference in reintubation rate was statistically signiifcant among infants with different gestational age (P<0.01). The infants with the gestation age≤28 weeks had a signiifcantly higher reintubation rate. According to whether the reintubation was performed, the infants were divided into success group and failure group. Compared to the success group, there were higher percentage of infants who had gestation age≤28 weeks, birth weight <1000 g and severe NRDS, needed high dose and repeated use of pulmo-nary surfactant and oxygen therapy, and had higher mortality in the failure group had (allP<0.05).ConclusionsThe INSURE technology can be effective in treatment of NRDS. Small gestational age, low birth weight, and severe NRDS are the risk factors for the failure of the INSURE technology.

Objective To investigate labial and lingual alveolar bone plate thickness of the mandibular anterior teeth by using cone-beam computed tomography (CBCT),in order to provide a reference for immediate implantation.Methods 132 individuals with normal occlusion were examined with CBCT,and their mandibular anterior teeth were analyzed.The labial and lingual alveolar bone plate thickness at the level of the apical,1/4 of the apical,1/2 of the middle and 1/4 of the cervical were measured.The differences of the thickness were compared among the side and gender.Results There were no significant differences between the left and right sides. Except on the 1/4 of the cervical,the thickness of lingual bone plate was thicker than that of labial bone plate.The frequency of the thickness of the labial bone larger than 2 mm on the 1/4 of the cervical of the center incisor,lateral incisor,and canine were 0,0 and 1.5 1%,respectively,and on 1/4 of the middle were 0.76%,1.44% and 3.79%,respectively.The lingual bone plate thickness showed significant difference between genders.Conclusion Reference values of alveolar bone plate thickness in anterior teeth of normal occlusion were established by using CBCT,which can provide clinical instruction for immediate implant.

[ABSTRACT]AIM:Toinvestigatecellapoptosisindiabeticfootulcersandtheeffectofadvancedglycosylation end products (AGEs) on apoptosis in human fibroblast cells.METHODS: Diabetic foot patients (n=18) and 18 age-matched non-diabetic controls were recruited .The clinical and biochemical features were compared by statistics methods . Skin biopsies were obtained from foot .Cleaved caspase-3 was measured by immunohistochemistry using the technique of streptavidin-biotin complex ( SABC ) staining.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling ( TUNEL) technique was used to detect apoptosis of the skin tissues .Human primary foreskin fibroblasts were isolated and cultured in the presence of 5.6 mmo/L glucose, 25 mmo/L glucose, fluctuant glucose ( changing the glucose from 5.6 mmo/L to 25 mmo/L every 8 h) or AGEs (150 mg/L, containing 5.6 mmo/L glucose).After 72 h treatment, Western blotting was used to determine the levels of the apoptotic protein cleaved-caspase-3.Other cells were trypsinized , washed with cold PBS and incubated with PI and Annexin V-FITC, then analyzed by flow cytometry to detect cell apoptosis .RE-SULTS:Diabetic patients had higher levels of fasting blood glucose (FBG), 2-hour postprandial blood glucose (2 h PBG) and glycosylated hemoglobin A1c (HbA1c), and longer wound duration.The protein level of cleaved caspase-3 was signifi-cantly higher in diabetic group , suggesting that apoptosis was increased in diabetic skin tissues .TUNEL analysis showed that apoptotic index was higher in diabetic group compared with that in non-diabetic group (8.4%±1.5% vs 3.8%± 0.8%) , which further confirmed that cell apoptosis was increased in diabetic foot tissues .In human fibroblasts , the levels of cleaved caspase-3 in normal group , sustained high glucose group , fluctuant high glucose group and AGEs group were 0.80 ±0.13, 1.22 ±0.18, 1.46 ±0.32 and 1.83 ±0.25, respectively.The apoptotic rates detected by flow cytometry were 2.43%±0.19%, 2.89%±0.51%, 3.99% ±0.24% and 6.83% ±0.36%, respectively.Both the level of cleaved caspase-3 and the apoptotic rate in AGEs group were higher than those in normal glucose group and sustained high glucose group .CONCLUSION:Increased apoptosis in diabetic foot ulcers is one of the most important reasons for im-paired wound healing .As compared to sustained high glucose and glucose fluctuations , AGEs induce greater apoptosis in human fibroblast cells .