Abstract:Objective To establish a rat model for lung injury induced by chest impact trauma. Methods Eight-four healthy male Sprague-Dawley rats were randomly divided into 7 groups (n=12) and were subjected to different chest impact energies (2.7,2.0,1.8,1.5,1.2,0.9 and 0 J) to induce bilateral lung injury using a self-designed device. The circulatory and respiratory functions were monitored before and after injury. The rats were sacrificed 120 min after chest impact. The morphological changes of the hearts and lungs were observed and graded pathologically. The wet/dry mass ratios of lungs and the protein contents in bronchoalveolar lavage fluid were determined. Results It was found that rats in the 1.2 J group developed bilateral lung contusion without substantial cardiac muscle trauma, and the animals had a low mortality. Conclusion Our self-designed device can satisfactorily produce lung injury model induced by chest impact trauma, and 1.2 J might be an ideal option for following studies.

Abstract:Objective To investigate the effect of hyperglycaemic memory on the local aldosterone system, reactive oxygen species (ROS) and expression of oncofetal fibronectin (oncofetal FN) mRNA in human mesangial cells (HMCs),and to further understand the role of local aldosterone system in the process. Methods In this study HMCs were divided into the following groups:normal glucose group (NG, 5 mmol/L D-glucose for 2 days),high glucose group (HG, 25 mmol/L D-glucose for 2 days), memory group (M, 25 mmol/L D-glucose for 2 days→5 mmol/L D-glucose for 4 days),memory+eplerenone group (MY,25 mmol/L D-glucose for 2 days→5 mmol/L D-glucose+10 μmol/L eplerenone for 4 days),normal glucose+eplerenone group (NY,5 mmol/L D-glucose for 2 days→5 mmol/L D-glucose+10 μmol/L eplerenone for 4 days),and persistent normal glucose group (SN, 5 mmol/L D-glucose for 6 days). ROS levels were tested by fluorescence microscope and fluorescence microplate reader. Aldosterone synthase (CYP11B2) protein expression was detected by Western blotting analysis. The mRNA expressions of 11β-hydroxysteroid dehydrogenase type 2,CYP11B2 and oncofetal FN were detected by RT-PCR. The expression and translocation of mineralocorticoid receptor (MR) was observed by laser scanning confocal microscope (LSCM). Aldosterone level in cell culture supernatant was detected by radioimmunoassay. Results (1)CYP11B2 mRNA and protein expression in group HG and in group M were all significantly increased, being 3.45, 2.09 and 3.14, 2.06 folds of those in group NG, respectively (all P<0.05). The aldosterone levels in HMCs culture supernatant were significantly increased in group HG and group M,being 2.01 and 1.81 folds of that in group NG, respectively (P<0.05). MR was activated and translocated from the cytosol to the nucleus in group HG and group M. Quantitative analysis showed that the ratios of cytosol/nucleus fluorescence intensity in group HG and group M were decreased by 30% and 21% compared with that in group NG, respectively (all P<0.05). (2)Oncofetal FN mRNA expression and ROS levels in group HG and group M were significantly increased,being 2.23, 1.99 and 2.16, 1.90 folds those of group NG, respectively (all P<0.05). Oncofetal FN mRNA expression and ROS levels in group MY were significantly decreased,being 35% and 51% of those in group M (all P<0.05). Conclusion HMCs have hyperglycaemic memory effect, which might be mediated by the local aldosterone system.

Abstract:Objective To investigate the effects of the guanine nucleotide exchange factor C3G (Crk SH3-domain-binding guanine-nucleotide-releasing factor) overexpression on the survival of cardiomyocytes and the underlying mechanisms.Methods H9C2 cardiomyocytes were transiently transfected with pCXN2-Flag or pCXN2-Flag-hC3G (overexpressing human C3G mRNA) plasmids, and then were exposed to hypoxia/reoxygenation (H/R) treatment. The cardiomyocytes were divided into blank group, empty plasmid group, C3G overexpression group, blank+H/R group, empty plasmid+H/R group, and C3G overexpression+H/R group. The expression of C3G mRNA was detected by RT-PCR, and the expression of C3G and p-ERK1/2 protein was detected by Western blotting analysis. The apoptosis rates of cardiomyocytes were analyzed by flow cytometry in each group, and the cell proliferation rates were analyzed by MTT.Results C3G overexpression increased cell proliferation rate and expression of C3G mRNA and protein, p-ERK1 protein(all P<0.01), and decreased apoptosis rate (P<0.05,P<0.01) compared to the blank and empty plasmid groups; the same was true for C3G overexpression+H/R group when compared to the blank+H/R and empty plasmid+H/R groups. Expression of p-ERK2 protein in C3G overexpression+H/R group was increased compared to blank+H/R and empty plasmid+H/R groups (P<0.01).Conclusion C3G overexpression can promote cell survival in the cardiomyocytes, which might be mediated by the increase of p-ERK1/2 protein expression and apoptosis inhibition.

Abstract:Objective To investigate whether COX-2/PGE2 can regulate vascular endothelial growth factor (VEGF) expression through activating Wnt/β-catenin signal pathway in human colorectal cancer cells. Methods PGE2 and (or) COX-2 selective inhibitor NS-398 was used to treat LoVo cells for 24 h, and then COX-2 and β-catenin protein expression was detected by Western blotting analysis. PGE2 and (or) β-catenin/tcf selective inhibitor FH-535 were used to treat LoVo cells for 24 h, and then the levels of VEGF was determined by ELISA. Routinely cultured LoVo cells were taken as control. Results Compared with the control group, PGE2 not only increased COX-2 protein expression, but also increased the levels of β-catenin in the total cell, cytosolic and nuclear proteins (being 3.8 folds, 2.7 folds, and 3.0 folds of the control, respectively, P<0.01). COX-2 selective inhibitor not only decreased COX-2 protein expression, but also decreased the levels of β-catenin in the total cell, cytosolic and nuclear proteins (being 0.3 folds, 0.3 folds, and 0.2 folds of the control, P<0.01). Compared with the control group, PGE2 increased VEGF expression in LoVo cells (being 1.6 folds of the control, P<0.01), and β-catenin/tcf selective inhibitor decreased VEGF expression in LoVo cells (being 0.68 folds of the control, P<0.01). Co-treatment of LoVo cells with β-catenin/tcf selective inhibitor and PGE2 showed no great effect on VEGF expression. Conclusion COX-2/PGE2 can increase β-catenin protein level, subsequently activate Wnt/β-catenin signal pathway and promote VEGF expression,which might be one of the mechanisms for COX-2/PGE2-induced angiogenesis in colon cancer.

Abstract:Objective To investigate the inhibitory effect of pioglitazone on inflammatory signaling pathways induced by human pancreatitis-associated ascitic fluid (PAAF) in human monocytic cell line THP-1. Methods PAAF was collected from patients with severe acute pancreatitis. The cultured THP-1 cells were divided into 4 groups: control group (C), PAAF group (A), PPARγ agonist pioglitazone group (P), and PPARγ antagonist GW9662 group (G). Cells in group A were stimulated with PAAF, those in group P were treated with 20 μmol/L PPARγ agonist pioglitazone 2 h after stimulation with PAAF,and those in group G were treated with PPARγ antagonist GW9662, pioglitazone (30 min later) and PAAF(2 h later) in turn.After incubation for 12 h, TNF-α and IL-6 mRNA expression was measured by real-time PCR, and Western blotting analysis was used to examine the expression of the phospho-p38-mitogen-activated protein kinase (MAPK), nuclear transcription factor (NF-κB) p65 levels, and IκB. Results Compared with group C, TNF-α and IL-6 mRNA expression in group A was increased, NF-κB and p38MAPK protein levels were increased, and IκB protein level was decreased (P<0.05). Compared with group A, TNF-α and IL-6 mRNA expression was decreased in group P, NF-κB and p38MAPK protein levels were down-regulated, and IκB protein level was increased (P<0.05). Compared with group P, TNF-α and IL-6 mRNA expression was increased in group G, NF-κB and p38MAPK protein levels were increased, and IκB protein level was decreased (P<0.05). Conclusion The results show that pioglitazone can inhibit PAAF-stimulated THP-1 inflammation by blocking p38MAPK and NF-κB signaling pathway.

Abstract:Objective To investigate the factors influencing the compliance of pre-exposure prophylaxis (PrEP) among high-risk groups of human immunodeficiency virus (HIV). MethodsA total of 352 subjects were selected into the cohort by non-probability sampling, including 153 men who have sex with men (MSM), 36 female sex workers (FSMs), and 163 fixed sex partners(PARs) of HIV carriers. Each subject was randomly given a medication regimen of either once daily or twice a week. The pre-exposure prophylaxis compliance was evaluated by observing the maintenance of the cohort and medication omission every 4 weeks from week 0 to week 28. Cochran-Mantel-Haenszel (CMH) test was employed to conduct univariate analysis and logistic regression was adopted for multiple variable analysis. Results It was found that 76.70% (270/352) of the subjects finished the 28-week-observation in this PrEP clinical trial among high-risk groups of HIV. The medication omission rate fluctuated around 35% during the follow-up, with the total compliance score being 82.12±27.48 and the median score being 95.24. Logistic regression suggested that the monthly income, presence of worries or difficulties during the trials, need for medical service, and the way of follow-up were the factors influencing the maintenance of cohort. The marital status, medication method, presence of worries or difficulties during the trial, and presence of adverse effect were the influencing factors of medication omission. The population type, number of sex partners, taking of other drugs at the same time, need of medical service during the trial, and way of follow-up were the factors impacting the overall compliance score. Conclusion The compliance of the present PrEP clinical trials among high-risk groups of HIV is high, and it has multiple influencing factors. Compliance management should be reinforced among MSM and FSW populations with greater motility. The cohort management model of follow-up by fixed specialists should be continued with more frequent follow-up; meanwhile, classified management should be implemented for individuals with different characters, so as to ensure a good compliance during the clinical trials.

Abstract:Objective To understand the growth and health care status of children under five years old in project counties of Chongqing and Sichuan, so as to provide evidence for establishing scientific and effective measures for health care of children. Methods The non-probability sampling method was used to select the children under five years old from 54 villages in 6 project counties in Chongqing and Sichuan. The parents were investigated by questionnaire; the height (length) and weight of children were examined; and the hemoglobin of 6-35 months old children was determined. Multivariable logistic regression analysis was conducted to examine the factors influencing the growth status of children under five years old. Results Totally 1 089 caretakers of children under five years old were investigated; 82.1% (894/1 089)of them had an educational background of primary school or middle school; and the median net per capita income of their family was 2 500 yuan (RMB) in 2009. For the youngest children in the families, the hospital delivery rate was 86.6% (933/1 077), the colostrums feeding rate was 96.4%(1 050/1 089), and the average of weaning months was (11.6±5.6) months. The prevalence of diarrhea of the children was 19.6%(213/1 089) within the past two weeks, and 264 children had cough (24.2%). A total of 1 197 children under five years old were investigated, the rates of stunting, underweight and wasting were 17.4%(200/1 152), 4.7%(55/1 173)and 2.2%(25/1 148), respectively. The underweight rate of boys was significantly higher than that of girls(5.9%\[38/645\] vs 3.2%\[17/528\]; χ2=4.638,P=0.031), and the wasting rate of boys was also significantly higher than that of girls(3.0%\[19/634\] vs 1.2%\[6/514\]; χ2=4.460,P=0.035).The detection rate of low hemoglobin among the 6-35 months old children was 38.4%(272/709). Multiple variable logistic regression analysis showed that male sex, born at home, and low birth weight might be the influence factors on the low growth status of children under five years old. Conclusion The caretakers of children under five years old in rural areas of Chongqing and Sichuan have a low cognitive level on the health care of children, and the children have developmental delay and malnutrition to various extents. Measures should be taken to improve child health care knowledge and health education.

Abstract:Objective To summarize our experience in using Smith-Robinson approach (SRA) for level C2/3 cervical spine surgery. Methods The clinical data of 36 patients receiving SRA for level C2/3 cervical spine surgery were retrospectively analyzed. The patients included 25 males and 11 females, with an average age of (47±13.5) years(range: 21-72 years). Thirty-five cases had Hangman fracture and one had traumatic disc herniation at C2/3. Combined injury included head injury (10 cases), soft tissue injury on the face (22 cases), fracture at lumbar spine (1 case), extremities fracture (6 cases) and injury of inner organs (2 cases). One patient had ankylosing spondylitis. Results C2/3 discetomy procedures were performed satisfactorily in all the 36 cases, with cages plus anterior plate fixation used in 34 cases and iliac bone plus anterior plate fixation in 2 cases. The patients were followed up for (9.1±2.8) months (ranging 6-32 months). One patient (2.4%) had choking and trouble in swallowing liquids, and the symptom disappeared 3 months later without any treatment. Neurological dysfunction was observed in 10 cases before operation, with the average JOA scope being (13.10±3.35) pre-operation and (15.70±1.49) post-operation, showing an improving rate of (74±23)%. Fusion was achieved in all cases during follow-up. Conclusion SRA can be applied at level C2/3 cervical spine surgery; the procedure is simple, with clear surgical field and less complication. Patients with large chin or extremely strong muscle may not be indicated for this approach.

Abstract:Objective To construct a three-dimensional(3D) anatomic model of the lateral ventricles for teaching microneurosurgical anatomy and clinical surgery planning.Methods The 3D model was constructed with a series of 176 MRIT2 head sections of a healthy male volunteer (36 years old) using software Matlab, Geomagic, Maya, etc. The 176 sections were subjected to management with software Matlab for image preprocessing and contour extracting; the acquired contours were subjected to software geomagic for 3D model constructing，3D-model and image connecting and other processings. Results The 3D model of the lateral ventricles and related structures were successfully constructed using more than 240,000 remodeled polygons, and the contour of the model consisted well with the images. The model had a even better visual effect after combining the 3D-model and image. The model could be viewed from any angle and could be rotated by 360 degrees in any plane using any structure as the focal point of rotation. Moreover, the model could be reduced or enlarged using the zoom function.Conclusion ConclusionThe constructed 3D model can be observed and studied repeatedly at any time, with no stress from surgery. The model may be used for interactive surgical exercises, paving a way for subsequent clinical practice.

Abstract:Objective To evaluate the advantages of microskin graft using acellular porcine skin for treatment of extensive deep burns by comparing with that using allogeneic skin. Methods A retrospective analysis was conducted on 70 severe burn patients who were treated in the Second Affiliated Hospital of Kunming Medical University during Jan.1999 to Jan. 2011. The patients were divided into the acellular porcine skin group and allogeneic skin group, each containing 35 patients. The survival rates of microskin grafts were determined at 4 weeks post-operation. Besides, the rejection of acellular porcine skin and allogeneic skin, changes of body temperature, white blood cell (WBC) count, lymphocyte and serum protein were observed at pre- and post-operation. Results (1) The survival rate was (71.5±6.6)% in acellular porcine skin group and (70.6±7.5)% in allogeneic skin group, with no significant difference found between the two groups (P＞0.05). (2) Acellular porcine skin group. At 3 days post-operation the acellular porcine skin was still attached to the wound, most of the skin was not discolorated, and small part of the skin became cinnamomeous. The acellular porcine skin was gradually separated from the auto-microskin at 3-4 weeks post operation, and there was small amount of exudates under the acellular porcine skin, which could be drained through a small cut. In the pressed area, there was still a small amount of exudates, but the acellular porcine skin was not dissolved and the microskin grafts survived and became confluent. (3) Allogeneic skin group. The allogeneic epidermal was rejected and was off from the wound at 3-14 days post transplantation, and at 10-30 days after transplantation the allogenic dermis became dry. During 25-60 days after transplantation, the allogenic dermis was completely stripped off, the microskin grafts became confluent, and the wound was healed. (4) The body temperature of the two groups was significantly descended after operation (P<0.01). The temperature of acellular porcine skin group was significantly lower than that of allogeneic skin group at 14 day post-operation (P<0.01). (5) The WBC count and percentage of neutrophils were also significantly decreased in both two groups after operation(P<0.05), and WBC counts in the acellular porcine skin group was significantly lower than those in the allogeneic skin group at 7 and 14 days after operation (P<0.05, P<0.01). The peripheral blood lymphocytes were also significantly increased after operation(P<0.01) and the lymphocytes in the acellular porcine skin group were significantly less than those in the allogeneic skin group at 14 days after operation (P<0.01). Blood biochemistry results showed no significant differences in the protein levels between the two groups (P＞0.05). Conclusion Microskin graft using acellular porcine skin, instead of allogeneic skin, for extensive burn patients can inhibit systematic inflammatory response, improve the nutrition condition, and reduce the using of allogeneic skin. Acellular porcine skin might be a suitable alternative for allogeneic skin.

Abstract:Objective To evaluate the safety and efficacy of radiofrequecy ablation for atrial fibrillation during minimally invasive mitral valve surgery via right thoracotomy. Methods From Jan. 2008 to Dec. 2011, 30 patients underwent radiofrequecy Maze Ⅲ procedure for atrial fibrillation during mini-invasive mitral valve surgery (study group). Another 30 patients with atrial fibrillation undergoing mitral valve surgery through median sternotomy without Maze procedure during the same period were taken as controls. The pre-treatment data of the patients were matchable between the two groups. The study group received mitral valve repair/replacement and radiofrequecy Maze Ⅲ procedure for atrial fibrillation. The operative outcome, postoperative complication and elimination rate of atrial fibrillation were compared between the two groups. Results No patient in the study group was transferred to median sternotomy during operation, and there was no reoperation, prolonged incubation, failure of important organs, hemoglobinuria or death. Compared with the control group, the study group had significantly longer mean circulation arrest time and cardiopulmonary bypass time, significantly reduced chest drainage and blood transfusion volume, and significantly shortened hospital study (P<0.05). The elimination rates of atrial fibrillation at immediately after operation,discharge and 6 months after operation were 96.7%, 66.7% and 73.3% in the study group, and 50%, 23.3% and 16.7% in the control group, respectively, with significant difference found between the two groups (P<0.01). Compared with the control group, better heart function recovery was achieved in the study group at 6 month after operation. Conclusion Radiofrequecy ablation for atrial fibrillation during minimally invasive mitral valve surgery via right thoracotomy is safe and effective. Importantly, it does not increase risks and complications of surgery. The early and middle term effects are satisfactory.

Abstract:Objective To observe the effect of serum of end stage renal disease patients undergoing maintenance hemodialysis (MHD) on the expression of ATP-binding cassette transporter A1 (ABCA1) in macrophages, and to study the interventional effects of different types of blood purification on the effect of serum. MethodsTotally 40 MHD patients were divided into hemodialysis (HD) group (n=23) and hemodiafiltration (HDF) group (n=17), and 23 healthy people were taken as controls. THP-1 macrophages were incubated with sera of subjects from each group. The serum lipid profiles and hs-CRP were measured by automatic biochemical analyzer. The serum levels of TNF-α, monocyte chemotactic protein 1 (MCP-1) and IL-6 were measured by ELISA. The mRNA expression of ABCA1 in macrophage was examined by RT-PCR. Western blotting analysis was used to measure protein expression of ABCA1,STAT3 and p-STAT3 in THP-1 macrophages. Results The levels of high-density lipoprotein cholesterol (HDL-c) and apolipoproteinAⅠ (ApoA-Ⅰ) in patients with end stage renal disease were significantly lower, and those of hs-CPR, TNF-α, MCP-1,and IL-6 were significantly higher than those in healthy group (P<0.05).The expressions of ABCA1, STAT3 and p-STAT3 in macrophages were not significantly different before treatment with HD and HDF, but the levels of ABCA1 in HD group and HDF group were significantly lower than those in healthy control group (P<0.05).The levels of ABCA1, STAT3 and p-STAT3 were not significantly changed before and after treatment in HD group. In contrast, the levels of ABCA1, STAT3 and p-STAT3 were significantly increased after treatment in HDF group(P<0.05). Conclusion HDF is more potent than HD in up-regulating ABCA1 in macrophages of MHD patients, enhancing its anti-inflammation and inducing cholesterol efflux, and the mechanism might be related to the activation of STAT3 signaling pathway.

Abstract:Objective To observe the pre-radiotherapy immunological state in patients with esophageal carcinoma, and to explore its association with the clinical pathological characters and prognosis. MethodsTotally 91 patients with pathologically confirmed esophageal cancer received radiotherapy. The proportions of T-lymphocyte subsets and natural killer (NK) cells in the peripheral blood were detected by the flow cytometry before radiotherapy; healthy population was taken as control. The correlation of immunological stress state and clinical pathological parameters were further evaluated in the patients. Univariate analysis (Kaplan-Meier) and multivariable analysis (Cox proportional hazards regression analysis) were used for survival analysis.Results The proportions of CD3+CD4+ (Th) T cells, CD8+CD28+ T cells and NK cells in pre-radiotherapy esophageal carcinoma patients were significantly lower than those in the healthy population (P<0.05). We also found that the proportion of CD3+CD8+ (Ts) T-lymphocytes was positively correlated with lymph node metastasis (P<0.05) and the proportion of NK cells was positively correlated with TNM stage (P<0.05). Univariate analysis showed that the patients with high proportions of Th or CD8+CD28+ T cells had a better survival (P<0.05). The multivariate analysis revealed that the proportion of Th cells before radiotherapy was an independent prognostic factor for patients with esophageal carcinoma. Conclusion The pre-radiotherapy proportion of Th cells in the peripheral blood is an independent prognostic factor for patients with esophageal carcinoma.

Abstract:Objective To establish a scoring formula of liver injury (SFLI) using matter-element analysis, so as to provide evidence for preoperative assessment and treatment of patients with liver cirrhosis diseases.MethodsThe preoperative serum biochemical indicators of liver cirrhosis patients were collected, including albumin (ALB), prealbumin (PA), total bilirubin (TBIL), serum creatine (SCr), alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyl transpeptidase (γ-GT), alkaline phosphatase (ALP), prothrombin time (PT), prothrombin time-international normalized ratio (PT-INR),activated partialthromboplastin time (APTT), and thrombin time (TT). The matter-element analysis was used to establish SFLI, and the results were compared with those of Child liver function system to create SFLI scores. Results An SFLI was successfully established. The scores of cirrhosis extents were expressed as the R value, which gradually decreased with the aggravation of cirrhosis. R=1 represented the normal liver tissue; 0.770≤R<1 represented early liver cirrhosis(compensatory cirrhosis), named SFLI Ⅰ; 0.712≤R<0.770 represented a state between compensatory cirrhosis and decompensated cirrhosis, named SFLI Ⅰ+; 0.629≤R<0.712 represented decompensated cirrhosis combined with ascites, named SFLI Ⅱ; and 0.401≤R<0.629 represented serious decompensated cirrhosis combined with ascites, different degrees of hepatic encephalopathy, and hepatic coma, named SFLI Ⅲ. Conclusion SFLI is a sensitive and accurate formula to demonstrate liver damage and liver function reserve during cirrhosis. It may help the preoperative assessment and treatment of the cirrhosis diseases in clinic.

Abstract:Objective To evaluate the effect of preparation process on the bonding strength of the nitrogen-doped TiO2-xNx films on brackets. MethodsRadio frequency magnetron sputtering was used to prepare nitrogen-doped TiO2-xNx films on the surface of stainless steel brackets. Post annealing was performed on TiO2-xNx films at 450℃ in nitrogen atmosphere for 2 h. The structures and surface morphologies of 3 types brackets, ordinary stainless steel brackets, nitrogen-doped TiO2-xNx films on brackets before annealing and the brackets after annealing, were evaluated by X-ray diffraction. The base plate of brackets was observed by scanning electron microscopy (SEM). The three types of brackets were also divided into 2 groups according to storing methods: distilled water at room temperature and thermal cycling. Sixty extracted human premolars (due to orthodontic demand) were bonded with 6 kinds of brackets using Jing-Jin enamel adhesive (n=10). All bonded specimens were tested on electronic universal testing machine to determine the bond strength. The data were analyzed by SPSS 17.0 software and the results were analyzed by one-way ANOVA. Results The TiO2-xNx thin films on brackets were in anatase phase, with the grain size being 30 nm. The base plates of 3 brackets were clear and well-organized, with no obvious differences. One-way ANOVA test showed that the bond strength of the brackets was significantly decreased after thermal cycling (P<0.01), and there were no significant differences among the 3 groups with the same storing methods (P>0.05). Conclusion The bonding strength of the nitrogen-doped TiO2-xNx films on brackets is not influenced by the preparation process, and can meet the demand of clinical orthodontic application.

Abstract:Objective To study the effects of morphine hydrochloride and temperature on the cephalopharyngeal skeleton of the larvae of Parasarcophaga crassipalpis in Shijiazhuan, Hebei, so as to provide evidence for forensic toxicologic entomology of sarcosaphagous flies and inference of the postmortem interval(PMI) in Shijiazhuang. MethodsAdult Parasarcophaga crassipalpis were collected. Four domestic rabbits were used in this experiment, with one receiving normal saline (control) and the others receiving morphine hydrochloride (0.5,1.0 and 2.0 folds of lethal doses) via ear vein injection. Then the rabbits were hit on the head and the muscle was marked with M0, M1, M2 and M3; then the larvae were transferred to the muscle sites at 24℃, 28℃ and 32℃. When larvae were laid, 10 of them were randomly sampled from each group at 12 h intervals until the beginning of the prepuparial stage. The larvae were collected every 12 h, fixed with boiling water, and stored as specimen. The scolex of the larvae was sparated and then the cephalopharyngeal skeleton was separated symmetrically, observed and photographed. Finally the indices, including the areas of different parts of cephalopharyngeal skeleton, sclerotized area, and average optical density were determined using digital image processing equipment. Results (1) The larvae of Parasarcophaga crassipalpis grew faster with the increase of morphine hydrochloride concentration and the temperature.(2)The color of the cephalophargngeal skeleton of the larvae changed from yellow to dark brown as time went by. Meanwhile, the degree and range of chitinazation were increased, accompanied by increase of the sclerotized area. The attached larval mouth (shaped like a dot) could be observed during the third age. (3) The areas of the larval mouth hook and the larval pharyngeal sclerite increased with time. Especially, remarkable changes were observed during the fold period, and the changes reached their maximum in the third age, and then remained basically unchanged. (4) The sclerotized areas and the average optical densities of the cephalopharyngeal skeleton kept an increasing tendency till the end of the third age. The periods needed for reaching the maximal optical density and sclerotized area were greatly longer than that needed for reaching the maximal area of the cephalopharyngeal skeleton. Conclusion The sclerotized area and the average optical density of the mouth hook and the pharyngeal sclerite of the larvae of Parasarcophaga crassipalpis keep increasing till the end of the third stage, and they are ideal indices for deducing PMI.

Abstract:Objective To investigate the genetic phenotypes of Pseudomonas aeruginosa isolates from naval divers, so as to obtain the epidemic character of Pseudomonas aeruginosa isolates in different naval divers. Methods A total of 64 Pseudomonas aeruginosa strains from naval divers were sequenced by multilocus sequence typing (MLST) using 7 house-keeping genes: acsA, aroE, guaA, mutL, nuoD, ppsA and trpE. The sequencing results were analyzed based on Pseudomonas aeruginosa international MLST database to obtain the allelic profiles and sequence types (STs). MLST data were analyzed by Bionumerics 4.0 (http://pubmlst.org/mlstanalyse) using LIAN, SplitsTree and eBURST. Results Data from MLST revealed a high diversity among the strains. Out of the 64 strains, 53 strains could be assigned to 19 STs and 11 could not be assigned. ST274 and ST260 were the dominant strains, with ST274 accounting for 18.75% (12/64) and ST260 accounting for 15.62% (10/64). Conclusion Pseudomonas aeruginosa strains in naval divers have high diversity and preponderant genotypes. MLST is of great significance for Pseudomonas aeruginosa genomic study and epidemiology surveillance.

Abstract:Objective To prepare a mitomycin C (MMC)-eluting stent for biliary tract, and to observe the drug delivery from the stent. MethodsThe mixed powder of MMC and polylactic glycolic acid (PLGA) was dissolved with their common solvent tetrahydrofuran (THF), with a drug concentration of 1%. The 6Fr biliary stent was soaked in the above solution for 10 min, and then was subjected to vacuum drying and was stored at room temperature. Then MMC-eluting stent was weighed and the MMC load was calculated. The MMC-eluting stents were soaked in 8 ml phosphate buffered saline (PBS, pH 7.4)and placed in a shaker at a constant temperature of 37℃ for 24 h soaking; then fresh PBS was changed every day for 30 days. The leaching solutions of the 1-30 days were subjected to chromatographic analysis to determine the concentrations of MMC. Results MMC load on each stent was (216.20±2.04) μg, with the load per unit area being (0.732±0.007) μg/mm2. MMC could be released from the stent surface in a sustainable manner. The elution concentration of MMC was (1.81±0.06) μg/ml on first day and (1.24±0.04) μg/ml on the second day; then it fluctuated within 0.61-0.84 μg/ml. The concentration of MMC began to decrease from the 21st day, and it was (0.51±0.01) μg/ml on the 30th day. Conclusion MMC-eluting biliary stents can be successfully prepared using polylactic acid as a drug carrier. In vitro study shows that the drug-eluting stents can sustainably and stably release MMC for over 30 days.

Abstract:The early symptoms and laboratory findings of pancreatic cancer are nonspecific and the outcome of victims are poor. Currently surgical resection is the only means for cure. When pancreatic cancer is clinically suspected, it is usually non-resectable due to metastasis. So prevention and early diagnosis are especially important for pancreatic cancer. The risk factors of pancreatic cancer include male sex, old age, long-term smoking, coffee consumption, alcohol abuse, diabetes, chronic pancreatitis, hereditary factors, obesity, non-familial hypercholesterolemia, high caloric intake, and environmental factors. Many studies have indicated that pancreatic cancer is closely related to K-ras mutations, and patients with high susceptibility have a higher probability of K-ras mutations. This paper reviews the value of K-ras mutation in diagnosis of pancreatic cancer, K-ras mutation in pre-cancerous lesions, and the relation between high-risk factors and K-ras mutations.

Abstract:Sleep disorder is the most common concomitant symptom following traumatic brain injury (TBI), with more than half of the patients representing insomnia, hypersomnia and circadian rhythm disorders. Long-term sleep disturbance can significantly affect the mental status, cognitive function and recovery of brain injury in TBI patients, which may increase the risk of TBI caused by traffic accidents. A series of neuroendocrine abnormalities caused by hypothalamic-pituitary-target gland axis dysfunction may be the main reason for sleep disorders. Currently, there is no uniform standard for the diagnosis of various types of sleep disorder after TBI. Hypnotic medications, whose curative effects on sleep disorder and complication have not been fully estimated yet, are the main treatment of sleep disorders after TBI. Further understanding of the mechanisms of various sleep disorders following different types of TBI can guide clinical therapies. Individualized treatment will play a positive role in rehabilitation of patients with sleep disorders following TBI.

Abstract:Bladder cancer is one of the most commonlyseen cancers of the urogenital system. Currently cystoscopy is still the most reliable method for diagnosis of bladder carcinoma. The related technologies have progressed greatly over the past several years. Based on the conventional cystoscopy, several strategies have been developed for diagnosis of bladder cancer, including fluorescence cystoscopy, narrowband imaging cystoscopy, optical coherence tomography cystoscopy, virtual cystoscopy, confocal microscopy, and so on. All these strategies have improved the diagnosis accuracy and can help the early diagnosis of bladder cancer, and thus improve the prognosis.

Abstract:Objective To discuss the incidence, influencing factors and management of various complications following cranioplasty. MethodsThe clinical data of 298 traumatic brain injury patients, who received surgical decompression, were retrospectively analyzed, and the risk factors of surgical complications following surgical decompression were analyzed. Results The patients included 193 males and 105 females, with a mean age of (38.23±10.86) years. Traffic accident (162/298, 54.36%) was the most common cause of the injuries, and surgical complications were found in 76 (25.50%) cases. Single factor analysis showed that age, admission GCS score, timing of surgery, hematoma location and hematoma volume were significantly associated with surgical complications. Logistic regression indicated that admission GCS score and hematoma volume were independent factors of surgical complications. Conclusion Complications are frequently seen in traumatic brain injury patients following decompressive craniectomy; patients with severe traumatic brain injury and great intracerebral hematoma are more likely to have complications. Early surgical intervention (<12 h) can help to reduce the complications.

Abstract:Objective To establish a method for determining the contents of ursolie acid and oleanolic acid in Qinggansanjie granules for quality control of the preparation. MethodsHigh-performance liquid chromatography with a diode-array detector (HPLC-DAD) was used for determination at the following conditions: The column was Chromsil-C18 (4.6 mm×250 mm, 5 μm), mobile phase was A: 0.1% (V/V) formic acid solution, B: methanol, A∶B = 12∶88. The flow rate was 1.0 ml/min, the temperature of column was 25℃, the detection wavelength was set at 210 nm, the injection volume was 50 μl, and the running time was 25 min. Results Ursolic acid and oleanolic acid could be separated on baseline, with the linear range being 6.1-97.6 μg/ml for ursolic acid and 6.4-102 μg/ml for oleanolic acid. The results of intra-day and inter-day precisions were both within 2% (n=3), and the average recovery was 95.4% (RSD 1.54%, n=6) and 99.2% (RSD 1.97%, n=6). Conclusion The present method is simple, rapid, accurate, and with satisfactory repeatability; it can be used for quality control of ursolic acid and oleanolic acid in Qinggansanjie granules.

Abstract:Objective To establish a high performance liquid chromatography approach for determining matrine and sophoridine contents in Ganlibao capsule. MethodsAgilent NH2 (4.6 mm×250 mm, 5 μm) column was used with 1% phosphoric acidethanolacetonitrile (V/V/V=102268) as the mobile phase. The flow rate was 0.8 ml/min, and the temperature was at 25℃.The detection wavelength was set at 210 nm. Results The calibration curve of matrine showed a good linearity in the range of 5.21-156.3 μg/ml (r=1) and that of sophoridine in the range of 5.29-158.7 μg/ml (r=0.999 9). The equations of linear regression of matrine and sophoridine were Y=9.586 1X+1.003 8 and Y=7.604 3X-5.920 1, respectively. The recovery was 98.73% (RSD=3.67%) for matrine and 100.10% (RSD=2.24%) for sophoridine. The matrine content of 15 batches of Ganlibao capsule was (30.13±0.19)-(44.70±0.17) mg/g and the sophoridine content was (21.60±0.25)-(52.07±0.11) mg/g. Conclusion The present method is simple, accurate, and reproducible, and it can be used for the quality control of Ganlibao capsule.

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