BACKGROUND:In recent years, endoplasmic reticulum stress-caused apoptosis plays a crucial role in ischemia impairment and has become the hotspot of studies addressing myocardial ischemia/reperfusion (I/R) injury. The lower limb ischemic preconditioning (LIP) has the obvious protective effect on the immature myocardium, but until now, no study reports whether LIP effects on endoplasmic reticulum stress apoptosis in immature myocardial cells. OBJECTIVE:To investigate the effect of LIP on endoplasmic reticulum stress and apoptosis.
METHODS:Langendorff-perfused isolated rabbit hearts were used in this study. Twenty-four immature rabbits were randomized into three groups. Control group:Isolated rabbit heart was only perfused with Krebs-Henseleit for 180 minutes. I/R group:Isolated rabbit heart was perfused 20 minutes, and then ischemia for 60 minutes fol owed by reperfusion 100 minutes. LIP group:Limbs were repeatedly obstructed 5 minutes and relaxed 5 minutes for three times, to establish Langendorff models, and then repeated the method of ischemia/reperfusion in I/R group. The myocardial apoptosis was assayed with TUNEL method. The expression of glucose-regulated protein 78, Bcl-2, Bax and Fas was detected with western blot analysis.
RESULTS AND CONCLUSION:Compared with I/R group, apoptosis rate was significantly lower, the expression of Bcl-2 was significantly higher, and the expression of glucose-regulated protein 78, Bax and Fas was significantly lower in LIP group. This study demonstrated that LIP regulates myocardial cellapoptosis through reducing the expression of endopasmic reticulum stress GRP78, Bax and Fas and increasing the expression of Bcl-2.

Objective To compare open versus closed reduction in treatment of children with displaced femoral neck fracture whose reduction is delayed by more than 24 hours.Methods We retrospectively reviewed 51 children who had been treated at our department for femoral neck fracture between May 2005 and May 2012 after their reduction had been delayed for more than 24 hours.They were 37 boys and 14 girls,1.7 to 15.5 years of age (average,9.1 years).According to Delbet's classification,27 cases belonged to type Ⅱ,18 to type Ⅲ,and 6 to type Ⅳ.Of them,18 underwent closed reduction and 33 open reduction.Their complications were recorded in detail.Their therapeutic results were evaluated according to Ratliff's criteria and compared between the 2 methods of reduction.Results The 51 children were followed up for an average of 36.7 months (range,from 17 to 61 months).Anatomic reduction was achieved in 6 cases in the closed reduction group (33.3％) and 29 cases in the open reduction group (87.9％),respectively.All fractures healed after an average of 10.9 weeks (range,from 9 to 13 weeks),with one exception of nonunion in the closed reduction group.The incidence of femoral head necrosis in the closed reduction group (27.8％,5/18) was significantly higher than that in the open reduction group (15.2％,5/33) (P ＜ 0.05).According to the Ratliff's evaluation at the final follow-ups,the good to excellent rate in the closed reduction group(66.7％,12/18) was significantly lower than that in the open reduction group (84.8％,28/33) (P ＜0.05).Coxa vara occurred in 3 children in the closed reduction group but in none in the open reduction group.Conclusion Open reduction with internal fixation may lead to better outcomes than closed reduction in the treatment of children with displaced femoral neck fracture whose reduction is delayed by more than 24 hours,probably because open reduction can result in better reduction quality.

Biopsy ; Child, Preschool ; Female ; Humans ; Infant ; Lung ; pathology ; Lung Diseases, Interstitial ; diagnosis ; pathology ; Male

Bronchi ; abnormalities ; pathology ; Bronchial Diseases ; complications ; congenital ; diagnosis ; pathology ; Bronchoscopy ; Child, Preschool ; Diagnosis, Differential ; Female ; Humans ; Infant ; Male ; Pneumonia ; etiology ; physiopathology ; Respiratory System Abnormalities ; complications ; diagnosis ; pathology ; Trachea ; abnormalities ; pathology ; Tracheal Stenosis ; etiology ; pathology

Child ; Child, Preschool ; Diagnostic Errors ; Female ; Humans ; Infant ; Infectious Mononucleosis ; diagnosis ; Male ; Suppuration ; diagnosis ; Tonsillitis ; diagnosis

Child ; China ; Communicable Diseases ; Communicable Diseases, Emerging ; Humans ; Multicenter Studies as Topic

Objective To develop an assay to determine Mycobacterium tuberculosis resistance to rifampin, isoniazid, ofloxacin and amikacin by pyrosequencing and evaluate the value of this method in clinical application. Methods Eighty-nine clinical isolates of Mycobacterium tuberculosis from tuberculosis patients were collected from Shanghai Pulmonary Hospital during 2008 to 2009. All strains were isolated from decontaminated sputum, cultured on Lowenstein-Jensen media and identified by traditional biochemical tests with standard methods. Ten Mycobacterium tuberculosis were selected from the strain bank of Shanghai Pulmonary Hospital. The optimal conditions of detection of rpoB, katG, gyrA and rrs gene by pyroseuencing were determined, using the 10 Mycobacterium tuberculosis strains whose drug susceptibility of Bactec 960 and sequence of rpoB, katG, gyrA, rrs gene were known. Then the drug susceptibility of 89 Mycobacterium tuberculosis clinical isolate strains were detected by pyrosequencing using this conditions and the results were compared with that of the Bactec 960 methods. Results The pyrosequencing program of sequence analysis was suitable for the detection of the mutations of rpoB and gyrA genes, and the program of single nucleotide polymorphism was suitable for katG and rrs genes. Among the 89 Mycobacterium tuberculosis clinical isolates,when using the drug susceptibility of Bactec 960 as the standard, the sensitivity of rifampin, isoniazid,ofloxacin and amikacin is 98.0%, 64. 1%, 79.5%, 78. 3% respectively, the specificity is 97.5%,100. 0%, 90. 0%, 100. 0% respectively, the accuracy is 97.8%, 77. 5%, 85.4%, 94. 4% respectively,tested by pyrosequencing. The results of the detection of resistance to rifampin, isoniazid, ofloxacin and amikacin in Mycobacterium tuberculosis using pyrosequencing technique were almost the same with that of Bactec 960, and Kappa ≥0. 7 in each detection. Conclusion Pyrosequencing is thus a rapid, accurate and high throughput method for detecting Mycobacterium tuberculosis resistance to these four drugs.

Objective Optimal detection of deep hypothermic circulatory arrest (DHCA)-induced early brain injury is important but the effective technique is still not available in the present.The relationship between diffusion wcighted imaging (DW1) and histopathological changes in DHCA-induced piglet brain injury model were analyzed.Methods Eighteen pigs underwent deep hypothermic circulatory arrest and were divided into three groups:group A (n =6) served as control,only underwent anesthesia and thoracotomy,without extracorporeal circulation.Group B (n =6) served as underwent deep hypothermic circulatory arrest at 20 ℃ for 120 min,survived for 1 day.Group C (n =6) served as underwent deep hypothermic circulatory arrest at 20 ℃ for 120 min,survived for 2 days.Diffusion-weighted imaging and histopathology were used to study the brain injury.Results There were hematoxylin-eosin (+) and terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (+) cells in cerebral cortex and hippocampus.Hyperintensity was most evident in the cerebral cortex in group B and group C with diffusion-weighted imaging,but it was not seen in hippocampus in both groups.There were 5 and 6 piglets can be seen hyperintensity with diffusion-weighted imaging sequence in group B and group C respectively.Hyperintensity was also seen in T2WI sequence in 3 and 5 piglets in group B and group C respectively.Conclusion The study demonstrates the feasibility of diffusion-weighted imaging on evaluation of brain injury after deep hypothermic circulatory arrest and cerebral cortex was selectively vulnerable to cell injury.It is superior to conventional imaging.

Objective To investigate the alteration of C/EBP α,C/EBP β and endoplasmic reticulum stress (ERS) related molecules (IRE1α and sXBP1) expression in pancreatic tissues in rats with hypertriglyceridemia related acute pancreatitis.Methods Ninety six Sprague Dawley (SD) rats were divided into 4 groups:control group,hypertriglyceridemia (HTG) group (n =24,fed with high fat diet for 2 weeks),AP group (n =24),HTG + AP group (n =24),and AP was induced by peritoneal injection of cerulein.The rats were sacrificed at 3,6,9,24 h after AP induction,respectively.The pathological changes of the pancreatic tissues were observed and scored by HE staining.Plasma levels of IL-1β,IL-6 and TNF-α were measured by ELISA.The expression of IRE1α,sXBP1,C/EBPoα,C/EBPβ mRNA were analyzed by real time PCR.The expressions of IRE1α,sXBP1,NF-kB,C/EBPα and C/EBPβ protein were determined by Western Blot.The expressions of C/EBPα and C/EBPβ proteins were also determined by immunohitochemistry.Results After two weeks of high fat diet,serum levels of triglyceride(TG) and total cholesterol (TC) in HTG group,HTG + AP group were much higher than those of the control group,and the difference was statistically significant (P ＜ 0.05).The pancreatic tissue injury was more severe in HTG + AP group,particularly at 9 h (P ＜ 0.05).And plasma IL-1β,IL-6 and TNF-α levels were also much higher in HTG + AP group when compared with that of AP group,the differences were all significant at 9 h (P=0.011;P=0.034;P =0.027).After AP induction,IRE1,sXBP1,C/EBP and C/EBPβ mRNA began to be up-regulated at 3 h,and IRE1 mRNA reached the highest level at 24 h,sXBP1 mRNA at 9 h,while C/EBP and C/EBPβ mRNA reached the highest level at 6 h.Compared with AP group,IRE1,sXBP1,C/EBP and C/EBPβ mRNA levels were much higher in HTG + AP group.In addition,as to IRE1 and sXBP1 mRNA,the difference was significant at 3,6,9,24 h,and C/EBP mRNA at 6,9,24 h,C/EBPβ mRNA at 6 and 9 h (P ＜ 0.05).After AP induction,IRE1α,sXBP1 and NF-kB proteins in the pancreatic tissue began to be up-regulated at 3 h,and all reached the highest level at 9 h.IRE1α,sXBP1 and NF-kB proteins were up-regulated more obviously in HTG + AP group,and the up-regulation in HTG + AP group was higher than that in AP group,and the high expressions of C/EBPα and C/EBPβ proteins could only be detected at 6 and 9 h in the HTG + AP group,while there was no expression detected in AP group.Conclusions C/EBPα,C/EBPβ,IRE1α and sXBP1 may be involved in the pathogenesis of HTG related AP,and IRE1α/sXBP1 pathway and C/EBPoα,C/EBPβ may mediate the pathologic injury and inflammation process of HTG related AP.

Objective To analyze and compare two potency assay methods of protamine sulfate.Methods Heparin titration method was according to foreign pharmacopoeia,while biological assay method was according to China pharmacopoeia(2015).Results The absorbance at the end-point was stable at 0.10 mg/mL and 0.15 mg/mL test solutions.The relative standard deviations(RSDs)of potency results were all less than 5%.Three test solutions and volumes of the titrant had a good linear relationship(R2 =1.000).The results of titration method were significantly related to those of biological assay(P=0.013),with similar RSD(P>0.05).However,the potency of titration method were significantly lower than those of biological assay (P=0.045).Conclusion Heparin titration method is good-using,convenient,and need to be applied widely at home.