Objective To observe the effects of Tongyang Huoxue Recipe on cytoskeleton proteinβ-tubulin of ischemia reperfusion injured sinoatrial node cell in rabbits;To discuss its mechanisms in the treatment of sick sinus syndrome. Methods Sinoatrial node cells were obtained from newborn rabbit. Oxygen and glucose were deprived to simulate ischemia and were restored to simulate reperfusion. Cells were divided into 5 groups. Tongyang Huoxue Recipe high-, medium-, low dose groups were given corresponding medicine (final concentrations of 100, 20, 10 μg/mL). The normal group and model group were given equal volume of culture medium. Enzyme mark instrument and laser scanning confocal microscopy were used to observe the sinoatrial node cell activity and cytoskeleton protein β-tubulin of each group. Results Living cells of model group decreased significantly compared with normal group (P<0.01), and cytoskeleton proteinβ-tubulin cleavage significantly. Living cells of Tongyang Huoxue Recipe high, medium and low dose group were significantly higher than those in the model group (P<0.05), and β-tubulin structure remained more complete than the model group. Conclusion Tongyang Huoxue Recipe can inhibit the injury induced by ischemia reperfusion of sinoatrial node cells;the possible mechanism of Tongyang Huoxue Recipe in treating sick sinus syndrome is protecting activity and cytoskeleton proteinβ-tubulin of sinoatrial node cells.

Objective To study the effects of different hypoxic training modes on the activities of respiratory chain complexes in the brain mitochondria of rats. Methods Forty healthy two-month-old male Wister rats were randomly divided into 5 groups: living low-training low(LoLo), living high-training high(HiHi), living high-training low (HiLo), living low-training high(LoHi), and living high-exercise high-training low(HiHiLo). Rats were forced to complete a 5-week endurance training program. All training sessions were performed at the same relative intensity under normoxic(P=632mmHg, simulating about 1500m altitude) or hypoxic conditions(P=493mmHg, simulating 3500m altitude) for each of the five groups, respectively. Mitochondria were isolated with differential centrifugation. Spectrophotometric analysis was applied to evaluate RCC (Ⅰ- Ⅲ) activities in brain mitochondria. Results Compared with the LoLo, brain RCC activities in C Ⅱ in rats from LoHi significantly decrease(P<0.05). Brain RCC activities in C Ⅲ in rats from HiHi, HiLo and LoHi significantly decrease and from HiHiLo significantly increase (P<0.01). Conclusion These findings suggest that among different altitude training modes, HiHiLo is the superior one in developing function of mitochondria respiratory chain of rat brain tissues.

ObjectiveTo observe the effects of catalpol, the effective component of Rehmanniae Radix, on atrionector cell apoptosis andβ-tubulin in vitro rabbit injured by simulated ischemia reperfusion;To explore the mechanism of treating sick sinus syndrome.MethodsAtrionector cells were collected from newborn rabbits. Cells were divided into 5 groups:normal group, model group, catalpol high, medium and low dose groups. Anoxia and aglycaemia were established to simulate ischemia. Atrionector cellular damage models were established by recovering the supply of oxygen and sugar. Normal control group and model group were given the same volume of culture medium, while catalpol high, medium and low dose groups were given medicine with relevant concentrations (100, 20, 10μg/mL, respectively). ELISA, FCM, laser scanning confocal microscopy were used to observe apoptosis rate andβ-tubulin of atrionector in each group.Results Apoptosis rate of the model group was obviously higher than the normal group (P<0.01), andβ-tubulin cleavage was obvious. Apoptosis rate in catalpol high, medium and low dose groups were significantly lower than that of the model group (P<0.01);β-tubulin structure were significantly more complete compared with the model group;the fluorescence intensity was significantly higher than that of model group (P<0.01).Conclusion Catalpol can inhibit atrionector cellular apoptosis caused by simulated ischemia reperfusion. Its protective effects on atrionectorβ-tubulin may be the mechanism of the treatment of Rehmanniae Radix for sick sinus syndrome.

Objective To observe the effects of ferulic acid on cell apoptosis andβ-tubulinin vitro of rabbit sinoatrial node injured by hypoxia and low glucose, and to explore its mechanism.Methods Model cells were established on sinoatrial node cells from newborn rabbit by performing deprivation of oxygen and glucose. Then the model cells were divided into 5 groups: a normal control group, a model group, ferulic acid high, medium and low dose group. The normal control group and the model group were treated with equal volume of culture medium, and the three volume ferulic acid groups were treated with ferulic acid of 100, 20, and 10μg/ml respectively. Flow cytometry, laser scanning confocal microscopy were used to observe the sinoatrial node apoptosis rate and cytoskeleton proteinβ-tubulin in each group.Results Apoptosis rate of the model group was obviously higher than the normal group (56.95% ± 11.99%vs. 31.45% ± 6.32%,P<0.01), whileβ-tubulin cleavage was significantly lower than the normal group (5.141 ± 0.218vs. 8.035 ± 0.762,P<0.01). Apoptosis rate of ferulic acid high, medium dose group were significantly lower than the model group (24.85% ± 6.34%, 26.70% ± 9.84%vs. 56.95% ± 11.99%,P<0.01),β-tubulin structure were significantly more complete compared with the model group (7.927 ± 0.357, 5.961 ± 0.351vs. 5.141 ± 0.218,P<0.01).Conclusions Ferulic acid can suppress apoptosis of sinoatrial node cells caused by low glucose and oxygen. Protecting the sinoatrial node cell skeleton protein ofβ-tubulin may be one of the mechanisms.

Objective To investigate the tumor diameter on the prognosis of patients with advanced esophageal squamous cell carcinoma(ESCC) after Ivor-Lewis surgical resection.Methods The clinical data of 254 patients with advanced ESCC who received Ivor-Lewis surgical resection at the Affiliated Hospital of Tianjin Medical University from January 2005 to December 2008 were retrospectively analyzed.All the patients were followed up via outpatient examination,telephone interview and correspondence till December 2013.Survival curve was drawn by the Kaplan-Meier method,and survival rate was analyzed using the Log-rank test.Receiver-operating-characteristic (ROC) curve analysis was used to determine the appropriate cutoff value of tumor size.Univariate and multivariate analysis were done using the chi-square test and COX regression model.Results Of 254 patients,223 patients were followed up for a median time of 30 months (range,3-108 months) with a follow-up rate of 87.80％ (223/254).The median total survival time was 27 months,and the 1,3,5-year overall survival rates were 72.7％,42.2％ and 31.3％,respectively.ROC analysis showed that the appropriate cutoff value of tumor diameter was 3.5 cm.The median survival time and 5-year survival rate were 36 months and 39.3％ in patients with tumor diameter ≤ 3.5 cm and 18 months and 25.4％ in patients with tumor diameter ＞ 3.5 cm,respectively,with a significant difference (x2 =9.494,P ＜ 0.05).The results of univariate analysis showed that the age,tumor diameter,depth of tumor invasion,lymph node metastasis and postoperative adjuvant therapy were related factors affecting the prognosis of patients with advanced ESCC after Ivor-Lewis surgical resection (x2=4.459,9.494,6.993,10.382,5.507,P ＜ 0.05).The results of multivariate analysis showed that tumor diameter ＞ 3.5 cm,lymph node metastasis and no postoperative adjuvant therapy were the independent factors affecting the prognosis of patients with advanced ESCC after Ivor-Lewis surgical resection (HR =1.631,1.681,0.677,95％ confidence interval:1.151-2.312,1.198-2.358,0.487-0.942,P ＜ 0.05).Of 159 patients without postoperative lymph node metastasis,median survival time and 5-year accumulated survival rate were 49 months and 46.4％ in patients with tumor diameter ≤ 3.5 cm and 23 months and 32.0％ in patients with tumor diameter ＞ 3.5 cm,respectively,with a significant difference (x2 =6.412,P ＜ 0.05).Conclusions The tumor diameter ＞ 3.5 cm,lymph node metastasis and no postoperative adjuvant therapy are the independent factors affecting the prognosis of patients with advanced ESCC after Ivor-Lewis surgical resection,meanwhile there is an assessed value of tumor diameter on the prognosis of patients without lymph node metastasis.

Objective To study the characteristics of liver injury induced by simvastatin combined with HRZ (Isoniazid,Rifampicin and Pyrazinamide) in SD rats.Methods Fifty-four 8-week-old SD rats were randomly divided into 3 groups:group A (control),group B (HRZ) and group C (simvastatin combined to HRZ),half of each group were male.We calculated the accurate dose respectively before those rats were given intragastrical administration of corresponding drugs.Six rats were killed in each group on 10th,20th and 40th day,respectively.Before this,blood was fastened from femoral of every rat that would be killed to test liver function,liver tissue slices were made in order to observe the pathological characteristic.Results Alanine amiotransferase of group C elevated in line with time and reached statistic difference on 40th day,furthermore,it was significantly higher than group A (P＜0.05).Total bilirubin and direct Bilirubin of group C were significantly higher than those of group A from the beginning to the end (P＜0.05),however,they declined rapidly on 10th day,this trend also had statistic difference (P＜0.05) At the end of this experiment,hepatic cords was disordered slightly,but swelling liver cells and vacuolar degeneration were observed,the nuleus of cell condensed.Soakage of monocytes,neutrophils,and lymphocytes occurred in the portal and lobule regions,or even spotty necrosis occasionally.Conclusion Cholestasis occurs at the early stage when simvastatin is combined with HRZ in SD rats,however,it has a rapidly degressive trend.In contrast,Alanine amiotransferase elevates,furthermore,pathological injury or even spotty necrosis can emerge in liver tissue slices.

Objective To investigate the incidence and risk factors of acute kidney injury (AKI). Methods The clinical data of 438 patients with severe pneumonia who were admitted in the intensive care unit in 2016 were retrospectively analyzed for analysis of AKI and assessment of risk factors. Results Among the 438 patients with severe pneumonia,the incidence of AKI was 53.42%. Within 72 hours after admission into the ICU, 86.75% of them developed AKI mainly from hypovolemic or septic shock(accounting for 80.34%). The detection rate of AKI by the standards of RIFLE and AKIN was lower than by the standards of KDIGO (45.89%,48.17%vs. 53.42%). Anemia ,shock /hypotension ,drugs ,RRT ,APPACHE II scores were the predictors of AKI. Conclusions The incidence of severe pneumonia complicated with AKI is high. Most of the patients develop it in 72 hours after admission into the ICU. Anemia,shock/hypotension,drugs,RRT,APPACHE II scores are its risk factors.

Objective To study the difference of image quality which based on different reconstruction in head dual‐energy computed tomography angiography (DECTA) scanning ,and evaluate the radiation dose which using different tube voltage in DEC‐TA .Methods An anthropomorphic head phantom with simulated arteries was used for DECTA scanning by different scan parame‐ters (tube voltage 80/140 kV and tube voltage 100/140 kV) ,the data was reconstructed with filtered back projection (FBP) and Si‐nogram Affirmed Iterative Reconstruction (SAFIRE) .The CT values with SD were measured and calculate to SNR and CNR ,the dose length product (DLP) was recorded and invert to effective dose .The image quality and radiation dose were evaluated in differ‐ent imaging condition .Results The image noise of SAFIRE reconstruction was lower than FBP reconstruction in same san parame‐ters .The increase of tube current‐time product from 100 mAs to 300 mAs ,it shows the characteristics of linear tendency .When it was on 260 mAs ,the image quality become stable .The difference between the two groups(80/140 kV ,100/140 kV)of SD of CT value by SAFIRE was significant (P< 0 .05) ,the difference of SNR and CNR between the two groups was not significant (P>0 .05) .The effective doses of group A (80/140 kV) was significantly lower than group B(100/140 kV) .Conclusion Using tube voltage of 80/140 kV combine with SAFIRE algorithm in head DECTA can ensure the image quality ,which can significantly reduce radiation dose .

Objective To investigate the effect on and mechansm by which prostaglandin E_1(PGE_1)protects liver functions after hepatectomy.Methods In this study,82 cases undergoing hepatectomy were divided randomly into control group with conventional therapy(41 cases),and PGE_1 treatment group(41 cases)treated with liposomal prostaglandin E_1 in addition to conventional therapy.Postoperative hospital days,urinary volume and abdominal drainage volume were observed.Pre-and postoperative liver functions were measured.Results Postoperative hospital days(median time 22 days)in PGE_1 treatment group were significantly shorter than those(median time 26 days)in control group.The postoperative levels of alanine transaminase,aspartic transaminase,total bilirubin and albumen in PGE_1 treatment group recovered to preoperative levels more quickly,than control group.Postoperative urinary volume in patients of PGE_1 treatment group was significantly more than that in control group,while abdominal drainage volume was markedly less,although there was no significant difference in prothrombin time between the two groups.Conclusion In patients undergoing hepatectomy,PGE_1 is very useful and safe to protect and improve hepatic function,decreasing the level of bilirubin,preventing ascites,formation shortening hospital days,without causing prolongation of prothrombin time remarkably.

Objective To construct a plasmid DNA encoding G glycoprotein of respiratory syncytial virus(RSV) and investigate the protective immune response against RSV infection. Methods Recombinant plasmid DNA of pcDNA3.1~G was constructed by standard RT-PCR based cloning procedure. The immunogenicity of recombinant G protein transiently expressed in HEK293 cells was detected by Western blot. BABL/c mice were intramuscularly immunized with pcDNA3.1~G. Samples of lung, sera, bronchoalveolar lavage fluid(BALF) were collected before and after RSV challenge; virus titer in lung was detected by viral titration; sections of paraffin embedding lung tissues were stained by haematoxylin and eosin(HE) for histological analyses; sera anti-RSV IgG levels were examined by ELISA; Th1/Th2 cytokine were detected by ELISA kit, the T lymphocyte subsets of BALF was determined by immunefluorescence staining followed by flow cytometry. Results Plasmid DNA of pcDNA3.1~G was successfully constructed. The expressed target protein possesses immunogenicity. After challenge, pcDNA3.1~G immunized mice presented relieved pathological changes in lung as well as reduced lung viral titers. The RSV specific IgG was detected in sera of immunized mice. There was significantly increased number of CD25~+CD4~+ T cells in mice BALF. Conclusion We constructed a pcDNA3.1~G plasmid DNA vaccination which can induce evident protective cellular immunity against RSV infection in mice with the increased number of CD25~+CD4~+ T cell subpopulation.