Objective To breed and identify steroid receptor coactivator-3 (SRC-3) knock-out mice. Methods Heterozygote mice were bred and reproduced. Wild genotype, heterozygote genotype, and homozygote genotype would appear in offsprings of parents. Genome DNA extracted from the murine tails was subjected to PCR for genotype identification. Male homozygote mice were selected to mate with the female heterozygote mice for acquiring homozygote baby mice according to Mendel law. Results Breeding and reproducing were successful and more heterozygote genotype mice were reproduced. Conclusion Appropriate methods for breeding, reproducing, and identifying are the effective way for acquiring SRC-3 knock-out mice from heterozygote mice.

BACKGROUND: Dynamic balance of endothelin (ET) and calcitonin gene-related peptide (CGRP) plays an important role in occurrence and development of cerebral infarction.OBJECTIVE: To observe the effects of thromboiytic therapy with batroxobin(DF-521) on the levels of plasma ET and CGRP after cerebral infarction.DESIGN: Randomized controlled study.SETTING: Fourth Department of Gereology, General Hospital of Guangzhou Military Area Command of Chinese PLA.MATERIALS: The experiment was performed at the Fourth Department of Gereology, General Hospital of Guangzhou Military Area Command of Chinese PLA. Totally 15 New Zealand rabbits aged from 18-20 months, of common grade and either sex, with the mean body mass of (3.5±0.5) kg were selected.METHODS: A total of 15 rabbits were established into cerebral infarction models with modified autogeneic thrombus method. One rat died when selecting models. Other 14 rats were randomly assigned into control group and treatment group. DF-521 was slowly injected into vein of ear edge of rats in the treatment group at hour 2 after embolism. Saline of the same volume was slowly injected into rats of the control group. Changes of concentrations of plasma ET and CGRP in the treatment group and control group were observed before and after cerebral infarction.MAIN OUTCOME MEASURES: Changes of ET and CGRP contents 1 hour before operation, 2 hours after operation (before drug injection) and 6 hours after operation (4 hours after drug injection).RESULTS: Of the 15 included rabbits, 1 rat died when establishing model, so 14 rabbits were involved in the result analysis. ET level significantly increased at hour 2 after acute cerebral infarction operation in the treatment group and control group (P ＜ 0.05, P ＜ 0.01), and then decreased at hour 6 after operation (P ＜ 0.01 ,P ＜ 0.01 ). The decrease in the treatment group was larger than that in the control group (P ＜ 0.01 ). The level of CGRP markedly reduced at hour 2 after acute cerebral infarction operation in the two groups, and than increased at hour 6 after operation. The increase in the treatment group was larger than that in the control group, but there was no statistical difference (P ＞ 0.05).CONCLUSION: Body with cerebral infarction during acute stage has selfregulative mechanism on dysequilibrium of ET and CGRP. Early-used thrombolytic therapy with DF-521 can markedly increase the ET level,which may be one of the protective mechanisms of cerebral ischemical reperfusion injury.

Objective To investigate the causer of, and the way to deal with the measurement of protecting from vascular compromise in free flap transplantation. Methods To retrospectively study the clinical data collected from the 89th Hospital of PLA including 305 cases of vascular compromise in free flap transplantation held on June, 1983 to December, 2006. Accoding to the different tissue flaps and different sites to be repaired, 11 factors of vascular compromise of free flap transplantation were to be investigated including operation design, the recipe of free flap, the variation of vascellum, the skill of recipe, the operation opportunity, the match of vascellum, the debride of vascellum, the vascellum angiotasis, the vascular anastomosis, the management of vascular articulo and the infection. Results Among the 305 cases of free flap transplantation, arterial compromise existed in 270 cases, 10 cases developed arterial compromise during operation, 6 cases were due to the injury of nutrient vessel, the flap survived after the revascularization. 4 cases obtained part necrosis of 10 cases, and the rotation flap and skingrafting covered the raw surface. The rest 213 cases survived and 47 cased failed. Another 35 cases of vein compromise were obtained. Among them, 5 cases survived, part necrosis of 10 cases, and 20 cases failed. Conclusion Vascular compromise is the factor of necrosis in free flap transplantation. It is of cardinal importance to timely and correctly treat the vascular compromise in free flap transplantation whether happened intra-or postoperatively. This is the key to access high successful rate. Preventive measures are even more beneficial than proper management after its occurrence. The venous return disorder was the main factor of free flap transplantation failure.

Objective: To investigate the changes of haemodynamics, pulmonary mechanics and blood gas in volume controlled and pressure controlled ventilatory patterns during one-lung ventilation (OLV). Methods: 20 patients with patent ductus arteriosus (PDA) underwent left thoracotomy PDA ligation with right OLV. The patients were divided into three groups: two-lung ventilation with volume controlled (TLV-VCV), one-lung ventilation with volume controlled (OLV-VCV), and one-lung ventilation with pressure controlled (OLV-PCV). After two-lung ventilation with VCV, one-lung ventilation was started by VCV and the ventilation mode was then switched to PCV. All measurements were made 25 min after initiation of the ventilation mode. The respiratory mechanics index was measured by side stream spirometry (SSS), including peak airway pressure (Ppeak), plateau pressure (Pplat), airway resistance (Raw), lung compliance (Cdyn) and inspiratory and expiratory minute ventilation (Mvi, Mve). Cardiac output (CO), stroke volume (SV), systemic vascular resistance (SVR), left ventricular ejectiontime (LVETi), and aortic blood flow acceleration (ACC) were also measured, by using the transesophageal Doppler (TED) monitor. Arterial blood gases was determined in every ventilation mode. Results: Ppeak、Pplat and Raw were significantly higher during OLV-VCV than that during TLV-VCV (P

0.05). However,compared with others segments, significantly high per centage of TA-SLN was absorbed in colon(P0.05). With the decrease of Na+ or existence of deoxycholate, T ween 80,SLS or DNP in the perfusion solution, the absorption rate increased rem arkably(P

BACKGROUND: Cervical sympathetic ganglia block accelerates the re covery of the homeostasis of organic nervous-endocrine-immune system, butit is still unclear whether it can suppress the imbalance of homeostasis in duced by post-traumatic stress disorder. OBJECTIVE: To observe the effect of cervical sympathetic ganglia blockon the mortality of mice with combined radiation and burn injury, andwhether it can become an easy and effective method to treat secondarydamage after serious trauma. DESIGN: A randomized grouping design, an animal controlled experiment. SETTING: Department of Anesthesiology, Guangzhou General Hospital, Guangzhou Military Area Command of Chinese PLA.MATERIALS: The experiments were carried out in the Institute of Combined Injury, the Third Military Medical University of Chinese PLA between February 2004 and July 2005. Totally 160 Kunming mice were randomly divided into control group (n=50) and cervical sympathetic ganglia block group (n=50). In the control group, the mice were only induced to models of combined radiation and bum injury, and treated with injection of 0.3 mL saline at cervical part. In the cervical sympathetic ganglia block group, the mice were induced to models of combined radiation and burn injury, and then treated with cervical sympathetic ganglia block, once a day for 14 days continuously.METHODS: Methods to induce injury in the animals: ① Radiation injury: The mice were given even radiation of 60Coγ ray (5 Gy) at a distance of 1.5 m to the whole body, the rate of absorptive dosage was (5.17-5.33) mGy/s. ② Burn injury: After the radiation injury, coagulated gasoline was smeared on the back and burnt for 8 s to induce degree Ⅲ burn injury of 15% of the total body surface, which was proved by the pathological section. Methods of cervical sympathetic ganglia block: Cervical sympathetic ganglia block was given bilaterally, and then the mice were injected with 0.2 mL lidocaine (5 g/L), and it was observed whether the symptoms similar to Horner syndrome (hyperemia of conjunctiva, drooping eyelid,blushing, smaller eyeslit) occurred or not at 5 minutes after injection.MAIN OUTCOME MEASURES: The mortality at 2, 5, 7, 10, 20 and30 days after injury and the changes of the numbers of red blood cells,white blood cells and blood platelet in peripheral blood at 7, 14 and 21 days after injury were observed in both groups. The effects of cervical sympathetic ganglia block on the levels of tumor necrosis factor-alpha (TNF-α),interleukin-1β (IL-1β) and interleukin-6 (IL-6) in serum at 3, 6 and 14days after combined radiation and burn injury were also observed.RESULTS: All the 160 mice were involved in the analysis of results without deletion. ① Compared with the control group, the mortalities at 5,7, 10, 15, 20 and 30 days in the cervical sympathetic ganglia block group were significantly decreased [control group: 8%, 22%, 32%, 54%, 74%,82%, 90%; cervical sympathetic ganglia block group: 8%, 14%, 16%, 22%,28%, 34%, 56%]. ② Compared with the control group, the numbers of red blood cells, white blood cells and blood platelets in peripheral blood at 7,14 and 21 days after injury in the cervical sympathetic ganglia block group were significantly increased [at 21 days: red blood cells: 23.21×1012 L-1, 14.58×1012 L-1; blood platelet: 16.87×1011 L-1, 12.57×1011 L-1; white blood cells: 20.65×109 L-1, 14.58×109 L-1]. ③ The levels of TNF-α, IL-1β andIL-6 in serum at 3, 6 and 14 days after injury in the cervical sympathetic ganglia block group were significantly decreased as compared with those in the control group [at 14 days: TNF-α: 189, 365 ng/L; IL-1β: 14, 23 ng/L;IL-6: 70, 132 ng/L].CONCLUSION: Cervical sympathetic ganglia block can significantly decrease the mortality of animals with combined radiation and burn injury,and it is an easy and effective method to treat serious trauma, and the mechanism may be realized through accelerating the recovery of hematopoietic function and suppressing the excessive inflammatory reaction.

Objective To establish a method to isolate,culture and identify cardiomyocytes from neonatal mouse,estimate the cardiomyocytes survival rate,and identify the purity of cardiomyocytes.Methods Neonatal C5 7 mouse heart(within 3 days)was di-gested using trypsin and collagenase.Cardiomyocytes survival rate was estimated by trypan blue staining,and cardiomyocytes purity was identified byα-actin immunofluorescence staining.Results Mouse cardiomyocytes could be successfully isolated and cultured using neonatal mouse within 3 days.Trypan blue staining showed cardiomyocytes survival rate was>95%,and cardiomyocytes pu-rity was more than 95% demonstrated byα-actin immunofluorescence staining.Conclusion Under the strict experimental condi-tions,mouse cardiomyocytes can be successfully isolated and cultured with high survival rate and high purity.

Objective To investigate the antitumor effect of tumor necrosis factor-related apoptosis-inducing ligand(TRAIL) gene transfection mediated by adenovirus into human pancreatic carcinoma cell line Panc-1,and the mechanisms involved in this effect.Methods TRAIL gene was transfected into pancreatic cancer cell line Panc-1 by an adenovirus vector(Ad-TRAIL).Level of TRAIL mRNA expression was determined using RT-PCR,and TRAIL protein synthesis was evaluated with Western blot.Cell-growth activities were determined by MTT assay.The bystander effect was observed by co-culturing the Panc-1 cells with and without the transfected TRAIL gene at different ratios.Apoptosis in pancreatic cancer cells was detected by flow cytometry.Proaspase-8 and procaspase-3 proteins were determined by Western blot.Results The stable overexpression of TRAIL was detected in Panc-1 cells transfected by Ad-TRAIL.Ad-TRAIL significantly inhibited cell viability of Panc-1 cells.Furthermore,co-culture of cancer cells transfected with TRAIL resulted in the nontransfected cell inhibition by bystander effect.Moreover,the percentage of apoptotic cells was significantly higher in the Ad-TRAIL-treatment group compared to the control groups(P

Objective To analyze the clinical experience of double lumen endobronchial tube intubation in 41 cases of whole lung lavage .Methods Forty‐one patients with whole lung lavage in our hospital from February 2010 to February 2015 were retro‐spectively analyzed .The effect of bronchoscopy and auscultation location method were explored after double‐lumen endobronchial intubation in whole‐lung lavage .Results Among 41 cases ,39 cases were successfully located by using the auscultation location ,and other 2 cases were successfully positioned by using the bronchoscopy position after repeatedly auscultation location method resulting in lung isolation failure .The catheter depth in 19 cases was adjusted after using bronchoscopy ,and then double lung was isolated well and lavage was successfully conducted .Conclusion The prerequisite for success of whole lung lavage is accurate position of double‐lumen endobronchial tube .Bronchoscopy look positioning has become the gold standard of double‐lumen endobronchial tube position .

ObjectiveTo investigate the morning blood pressure surge(MBPS) and its relationship with 24-hour blood pressure variability and anti-hypertensive drugs in middle-aged and elderly hypertensive patients.Methods A total of 521 middle-age and elderly men were surveyed with ambulatory blood pressure monitoring and ambulatory electrocardiograms recordings as well as questionnaire investigation from January 2009 to December 2010.Subjects were divided into MBPS positive group and MBPS negative group according to the level of MBPS[ ＞35 mm Hg (1 mm Hg=0.133 kPa) or ≤35 mm Hg].ResultsIn all the cases,the prevalence of MBPS was 19.4％,of which the elderly and very elderly had higher prevalences ( 18.9％ and 21.8％,respectively) than the middle-aged (5.6％,both P ＜0.01 ).Significant differences could be found in age [ ( 81.6 ± 6.4 ) years vs ( 78.7 ± 9.7 ) years ],day mean systolic blood pressure [ ( 132.8 ± 13.3) mm Hg vs ( 128.8 ±13.3) mm Hg],fasting blood glucose [ (5.96 ± 1.59) mmol/L vs (5.68 ± 1.22) mmol/L] and 24-hour blood pressure variability between the two groups ( all P ＜ 0.05 ).Significant difference could be observed in the prevalence of MBPS between the diuretics-taking group and non-taking group (27.4％ vs 17.6％,P ＜0.05 ).ConclusionsThe elderly hypertensive patients are prone to appear MBPS phenomenon.Fasting blood glucose level,24-hour blood pressure variability may be associated with MBPS.Diuretic antihypertensive treatment may not be conductive for MBPS control.