Objective Previous study showed that the histopathological basis of visual function damage caused by optical nerve injury is apoptosis of retinal ganglion cells(RGCs). This procedure is regulated by P53, bax and caspase 3 genes. Present study aimed to observe the expression of bax, P53 and caspase 3 mRNA in RGCs after traumatic optic nerve damage in the rats by SYBR green I fluorescence quantitative PCR method. Methods The animal model of optic nerve injury was established in the right eyes of 56 adult Wistar rats by a fluid percussion brain injury device (FPI) . Animal were killed on days 1, 3, 5, 7, 9, 14, 28 days separately after injury. Other 16 Wistar rats were divided into normal control group and sham operation group. The total RNA was isolated from rat fresh retina tissue by Trizol method and was treated by reverse transcription to cDNA using 01igo(dt) 18 as primer and then amplified. The target fragments of bax, P53 and caspase 3 cDNA were linked with carrier pTZ57 R/T to construct recombined plasmids which were transformated to E. Coli DH5α by T/A clone method. Recombined plasmids were extracted with alkaline lysis method and the plasmids were selected in white colonies by ampicillin screening, EcoR I restrictive enzyme analysis, and their specificity was evaluated using DNA sequencing. The standard curves were created by plasmid DNA and the precise expression level of target genes in samples were determined using software. The results were expressed as the ratios of target gene mRNA to GAPDH mRNA. Results The standard curve drawn by pTZ57R/T and target gene presented a good linear tendency with the higher sensitivity and specificity. The expression of P53 and bax mRNA began to increase on the third day after the injury of optic nerve and peaked on the fifth day and started to decline on the seventh day. The expression of caspase 3 mRNA increased from the fifth day through the ninth days after injury and declined on the fourteenth day. The significant differences were found in the expression of P53, bax and caspase 3 between model group and control group (P ＜ 0. 05) . Conclusion The pro-apoptotic protein P53, bax and caspase 3 play an important role in RGCs apoptosis.

Objective To develop a method for rapid and accurate detection and identification of bacterial pathogens directly from body fluid specimens and to evaluate its application in clinical laboratory.Methods Bacteria DNA was extracted from 205 body fluid specimens with column-based kit,and the high variable V1 and V3 regions of bacterial 16S rRNA gene were amplified with broad-range primers.Amplicons were analyzed by pyrosequencing and the generated sequences were searched in the bacterial identification database.Traditional culture-biochemical method was also used for these specimens and the results were taken as the golden standard.SPSS 11.0 was used to calculate the sensitivity,specificity,false positive/negative rate,positive/negative predictive value and positive/negative likelihood rate of pyrosequencing method.Results The positive rate of bacteria culture was 39.5％ (81/205),among which 71 were infected with single bacterium,and 10 were infected with two species of bacteria.Compared with the culture identification results,pyrosequencing had a 100.0％ (71/71) concordance when applied to detect and identify bacterial pathogens from specimens with single specie bacterium infected.To specimens with two species bacteria infected,7 out of 10 specimens were in concordance with the culture identification results.Besides,pyrosequencing detected 10 positive specimens and identified bacterial pathogens infected in the 124 culture-negative specimens.Taken bacteria culture as the standard method,the sensitivity of pyrosequencing for identifying bacterial pathogen in body fluid was 100.0％,and with a specificity of 91.9％,the false positive rate was 8.1％,the false negative rate was 0.0％,the positive predictive value was 89.0％,the negative predictive value was 100.0％,and the positive and the negative likelihood rate were 12.4 and 0,respectively.Conclusion Pyrosequencing can be used to detect and identify bacterial pathogens directly from body fluid specimens with the advantages of rapidity,high sensitivity,high accuracy and high throughput.

ObjectiveToexplore the appearance ofmusclecrushinjuryatquantitative ultrasonographic elastography by supersonic shear imaging (SSI).MethodsThe animal experiment was done using a special balloon cuff device to create left hind leg crush injury with a force of 18.6 kPa.Twentythree New Zealand rabbits had crush injury of extremity and survived for 72 hours.SSI quantitative elastography was performed in crushed and no-crushed regions of each rabbit hind leg.Quantitative lesion elasticity was measured in terms of the Young modulus (in kilopascals) at 30 min,2 h,6 h,24 h and 72 h after the release of the crushing pressure.A receiver operating characteristic (ROC) curve analysis was used to assess diagnostic performance.ResultsThe area of crushed region in left hind leg accounted for 2.6％ -3.0％ of body surface area in 23 rabbits.The crushed regions exhibited maximum elasticity values of (19.51 ± 6.74)kPa,(21.47 ± 5.54) Pa,(11.36 ± 5.35)kPa,(15.09 ± 3.31)kPa and (13.72 ± 3.74) kPa,and mean elasticity values of (12.44 ± 3.77)kPa,(13.20 ± 3.60)kPa,(6.80±2.86)kPa,(10.04 ± 2.95)kPa and (6.94 ± 0.97)kPa at 30 min,2 h,6 h,24 h and 72 h after the release of the crushing pressure.Comparing with those of no-crushed regions,they were higher obviously (P＜0.001).ROC curves showed that extremity crush injury was diagnosed by using elasticity value,and the greater the elasticity value,the greater the diagnostic value.Conclusions SSI provides quantitative elasticity measurements,thus adding complementary information that potentially could help in crush injury characterization with conventional ultrasonography.

Objective To evaluate the efifcacy and safety of hemostatics of injected gelatin matrix (HIGM) under the guidance of contrast-enhanced ultrasound (CEUS) for treating splenic trauma in canine model. Methods A total of 24 commercial hybrid dogs underwent celiotomy with creation of uniformly blunt splenic trauma lesion of 4.0 cm×4.0 cm×2.5 cm (length, width and depth, respectively) by hemostatic clamp. Subjects were prospectively randomized into two groups. The treatment group was treated with HIGM under the guidance of CEUS and the positive control group received thrombin solution. Conventional ultrasound and CEUS were performed to record the ascites and the splenic lesion areas at 1st, 3rd, 7th, 14th and 21st day. The ifne needle biopsy and splenectomy were performed for histopathologic examination. The weight, free intraperitoneal lfuid and injury site were compared with t test between HIGM and postive group. Results All animals in two groups survived. All dogs stopped hemorrhage after injection of HIGM under CEUS guidance. The area of injury site was (12.91±0.89) cm2, (4.45±0.75) cm2 and (1.38±0.23) cm2 at 1st, 3rd and 7th day and splenic lesions were not found at 14th and 21st day in all dogs (n=12) of HIGM group. The splenic lesion was (16.74±0.91) cm2, (11.26±0.99) cm2, (8.02±0.82) cm2 and (1.58±0.36) cm2 in the postive group at 1st, 3rd, 7th and 14th day and splenic lesions were not found at 21st day in all dogs (n=12). At 7th and 14th day post-injection, lesion areas were statistically significant between two groups (t=27.162, P=0.008;t=15.129, P=0.001). Free intraperitoneal lfuid was (0.91±0.05) cm at 1st day detected by conventional ultrasound and free intraperitoneal fluid was not found at 3rd, 7th, 14th and 21st day in all dogs (n=12) of HIGM group. The free intraperitoneal fluid in thepositive group was (1.96±0.17) cm, (1.30±0.11) cm and (0.81±0.12) cm at 1st, 3rd and 7th day and free intraperitoneal lfuid was not found at 14th and 21st day in all dogs (n=12). At 1st, 3rd and 7th day post-injection, free intraperatitoneal lfuid was statistically significant between two groups (t=20.934, P=0.003; t=41.310, P=0.000; t=22.520, P=0.000). Histopathological examination showed that there was no foreign body and foreign body granuloma and the structure of red pulp was recovered at 7th, 14th and 21st day. Gross anatomy showed that the splenic injury site was recovered completely without complications. Conclusion This study explored the value of HIGM for splenic trauma and provided a preliminary experimental evidence for clinical treatment.

Objective To design a new type of biliary stent to be used in the common bile duct probe and evaluate its biocompatibility, safety and effectiveness in a canine model.Methods Magnesium alloy (AZ 31B) was used to make biliary stents.A canine model of acute obstructive jaundice was established by ligating the distal end of the common bile duct.These dogs were divided into two groups with either placement of magnesium stent or not.The incidence of bile duct leakage and survival were evaluated.Meanwhile, we determined the concentration of magnesium in various body fluids and organs, liver function test, and ultrasonic and histological studies.Results The concentrations of magnesium in the blood, feces, liver, heart, brain and lungs were similar between the two groups.Bile leakage rate in experimental group and control group was 0％ and 16.7％, respectively.The mortality was 16.7％ and 33.3％.There were no significant differences in ALP and TBIL between experimental group and control group (P>0.05) before building and bile duct exploration.There were significant differences in ALP andTBIL between the two groups one and three months after bile duct probe (P<0.05).Biliary tract specimens showed obviously anastomotic stenosis performances and expansion performances of extrahepatic biliary in the control group, but in experimental group such changes were not observed.Ultrasonography showed that the patency of biliary anastomosis was good and there was no anastomotic biliary stricture in experimental group.However, progressive stenosis and expansion performances of extrahepatic biliary occurred in control group.There were fewer collagen fibers but more muscle fibers in experimental group than in control group.Conclusion Biliary stent made of magnesium alloy is of good biocompatibility and has the potential to prevent the postoperative stenosis in the common bile duct probe.

Objective:To investigate the role and mechanism of leptin on dendritic cells by NLRP3 inflammasome.Methods: BMDCs were induced in vitro,leptin with scalar doses was cocultured with BMDCs,IL-1β and IL-18 mRNA expression and protein secretion level were measured by q-RT-PCR and ELISA respectively.Caspase-1 activity or ROS synthesis were tested with FLICA kit or ROS detection assay kit on flow cytometry.IL-1β or IL-18 were detected after caspase-1 was inhibited by Ac-YVAD-cmk or NLRP3 was interfered by siRNA or ROS inhibitor DPI or KCL were added.Results: Leptin promoted secretion of IL-1β and IL-18.Leptin up-regulated NLRP3 and activted caspase-1 to secret proinflammtory cytokine,which K+ efflux took part in.Conclusion: Leptin promotes secretion of IL-1β and IL-18 by activating NLRP3 inflammasome,and K+ efflux takes part in this,which hints us that leptin may be an activator of NLRP3 inflammasome.

Objective To study feasibility of combined haemostatic percutaneous injection therapy guided by contrast-enhanced ultrasonography (CEUS) in treatment of renal injuries. Methods Eighteen New Zealand rabbits were inflicted with kidney injury imitating grades Ⅲ-Ⅳ blunt injuries. The animals were randomly and equally divided into three groups, Group A ( treated with hemocoagulase),Group B ( treated with hemocoagulase and Alpha-cyanoacrylate) and Group C ( control group, given normal saline). The hemostatic time, hemostatic effect, and perirenal hematoma were observed. Results A perirenal hematoma was observed one hour after treatment. The perirenal fluid thickness was (0.200 ±0.012) cm in Group A, (0.050 ±0.002) cm in Group B and (0.400 ±0.009) cm in Group C, with statistical significance between two test groups and Group C (P ＜ 0.05 ). At days 7 and 14 following treatment, lesion length and cross section was ( 1. 107 ±0. 143) cm and (0.433 ±0. 163) cm in Group A, (0.567 ±0.082) cm and (0. 160 ±0. 078) cm in Group B, and (0.980 ±0. 203) cm and (0.686 ± 0. 157) cm in Group C. There was statistical significance between the test groups (Groups A and B) and Group C (P＜0. 01) at day 14. The lesion size in Group A was lager than that in Group B (P ＜ 0.01 ). One month after treatment, a slight nephrohydrosis occurred in Group B. Conclusions Either injection of simple hemocoagulase or combined use of hemocoagulase and Alpha-cyanoacrylate guided by CEUS can attain positive hemostatic effect, but the latter one is more rapid and reliable.

Objective To approach the diagnostic value of contrast-enhanced ultrasonography(CEUS)for the detection of traumatic laceration of pancreas. Methods Sixty cases of pancreatic traumatic model were made in twelve healthy swines after the animals were anesthetized and laparotomized. Then the conventional ultrasonography(US) and CEUS were performed in each case to diagnose the traumatic region,immediately. The results were compared with surgical findings. Results Among sixty injuries,the detection rate of conventional ultrasonography was 66. 7%,the detection rate of CEUS was 88.3%. Conclusions CEUS shows higher detection rate than conventional US in diagnosing pancreatic laceration,and it also can improve the diagnostic value of ultrasound for the detection of pancreatic laceration.

Objective To study haemostatic percutaneous injection therapy for the management of vascular damage in patients with renal injuries guided by contrast-enhanced ultrasonography(CEUS).Methods Which of 56 patients with renal trauma were diagnosis by CEUS,37 cases with grades Ⅱ-Ⅳ renal injuries were brought into our study.According to wound degree and accompanying active bleeding,they were divided into experiment group (percutaneous injection hemostatic treatment)and control group(conservative treatment).Results Thirty-seven renal trauma manifest low perfusion in lesions by CEUS,and the contrast agent could be seen overflow to renal pelvi and the location of capsule in 13 patients.The patients were divided into experiment group(17 cases)and control group(20 cases).The color of hematuria of 9 patients in experiment group became gradually light at 30 mins after treatment.and the color of 7 cases become normal,and hematuria of the only one was iterative appear.The color of hematuria of 9 patients in control group became gradually light in 24-72 hours,others' hematuria became gradually light in 5-14 days.The time of color of hematuria become light of the former was shorter than those of the latter(P<0.05).Reexamination by ultrasound and renal function and urine routine at 1,3 and 6 months after treatment,the results of all patients indicated normal.Conclusions Haemostatic percutaneous injection therapy for renal trauma guided by contrast-enhanced ultrasonography has very obvious hemostatic efficacy.Its advantages included may be used for effective,minimally invasive control of renal injuries(grades Ⅱ-Ⅳ),and can be a feasible management of active bleeding at bedside.

Objective: To explore the effect of moxibustion at Shenque (CV 8) on myocardial structure and function in exercise-induced fatigue rats. Methods: A 12-week treadmill running training was performed to create an exercise-induced fatigue rat model. Sixty eligible male specific-pathogen-free grade Sprague-Dawley rats were randomly divided into a blank group, a control group, a model group, a non-meridian non-acupoint group, a Zusanli (ST 36) group and a Shenque (CV 8) group, with 10 rats in each group. Rats in the blank group did not receive treadmill running training or moxibustion. Rats in the control group did not receive treadmill running training but received mild moxibustion at Shenque (CV 8). Rats in the model group received treadmill running training but no moxibustion. Rats in the non-meridian non-acupoint group, the Zusanli (ST 36) group and the Shenque (CV 8) group received moxibustion at the non-meridian non-acupoint points, Zusanli (ST 36) or Shenque (CV 8) immediately after each treadmill running training, 15 min each time, once a day for 5 consecutive days a week at a 2-day interval, 60 times of moxibustion in total. Left ventricular end-diastolic diameter (LVEDd), left ventricular end-systolic diameter (LVESd), left ventricular diastolic volume (LVDv), left ventricular systolic volume (LVSv), ejection fraction (EF), stroke volume (SV), early diastolic peak flow velocity of mitral valve (E) and late diastolic peak flow velocity of mitral valve (A) of each group before and after the last treadmill running training were measured. Blood was collected 6 h after the last treadmill running training, and serum C-reactive protein (CRP), myoglobin (Mb), creatine kinase-myocardial band (CK-MB), cardiac troponin I (cTnI) and cardiac troponin T (cTnT) levels were detected. Finally, the heart was separated, the heart mass (HM) was measured, the cTnT level of the myocardial tissue was detected, the ultrastructural changes of the left ventricular myocardium were observed by transmission electron microscope, the left ventricular fraction shortening (LVFS), E/A and heart mass index (HMI) were calculated. Results: Compared with the same group before treatment, the rat cardiac LVEDd, LVESd, LVDv, LVSv, SV, E and A were significantly increased (all P<0.01), and the rat LVFS, E/A and EF were significantly decreased (all P<0.01) in the model group and the non-meridian non-acupoint group after treatment; the rat cardiac SV, LVDv, LVSv, E and A were all increased (all P<0.01), while E/A and EF were decreased (all P<0.01) in the Zusanli (ST 36) group after treatment; the rat cardiac LVDv, E and A were significantly increased (P<0.01 or P<0.05), and E/A was significantly decreased (P<0.01) in the Shenque (CV 8) group after treatment. After treatment, compared with the blank group, the rat cardiac LVEDd, LVESd, SV, LVDv, LVSv, E, A, the serum CRP, Mb, CK-MB, cTnI, cTnT and HMI, and the myocardial cTnT were increased (all P<0.01), and the LVFS, E/A and EF were all reduced (all P<0.01) in the model group; compared with the model group and the non-meridian non-acupoint group, rats in the Zusanli (ST 36) group and the Shenque (CV 8) group showed decreased LVEDd, LVESd, SV, LVDv, LVSv, E, A, serum CRP, Mb, CK-MB, cTnI, cTnT and HMI, and myocardial cTnT (P<0.01 or P<0.05), along with increased LVFS, E/A and EF (all P<0.01); compared with the Zusanli (ST 36) group, Mb and A of the Shenque (CV 8) group were decreased (both P<0.01), while both E/A and EF were increased (P<0.01, P<0.05). Transmission electron microscopy examination showed that myofibrils in the blank group and the control group were neatly arranged with clear light and dark bands; the model group and the non-meridian non-acupoint group showed different degrees of myofibril disintegration and breakage, increased and aggregated mitochondria of different sizes, and increased electron density. The myofibrils in the Shenque (CV 8) group and Zusanli (ST 36) group were arranged neatly with clear light and dark bands, and compensatory hyperplasia of mitochondria. Conclusion: Moxibustion at Shenque (CV 8) and Zusanli (ST 36) both can effectively improve the occurrence of myocardial remodeling in exercise-induced fatigue rats, and the effect of moxibustion at Shenque (CV 8) is better in improving cardiac function.