After human umbilical vein endothelial cells (HUVECs) were incubated with various concentrations of exenatide for 24,48,and 72 h under the conditions of5.5 and 33 mmol/L glucose,the cell viability was detected by MTF assay.The apoptosis rate of endothelial cells was measured by flow cytometry.Protein kinase B (Akt) phosphorylation,Bcl-2,and Bax protein expression were detected by Western blotting.The results showed that the cell viability was decreased after HUVECs were incubated with high glucose for48 and 72 h (P＜0.01).Treatment with 1,10,and 100 nmol/L exenatide for48 h significantly increased the cell viability in the presence of high glucose,in a dose-dependent manner (P＜0.01).High glucose inhibited Akt phosphorylation and Bcl-2 protein expression,stimulated Bax protein expression,with decreased Bcl-2/Bax ratio and increased apoptosis ratio of cells.After treatment with exenatide,the cell apoptosis reduced,Akt phosphorylation and Bcl-2 protein expression increased,and Bax protein expression decreased,with increased Bcl-2/Bax ratio (P ＜ 0.01).These effects of exenatide on endothelial cell were abrogated by an inhibitor of phosphotylinosital 3 kinase (PI3K) LY294002 (P ＜ 0.01),suggesting that the exenatide may regulate Bcl-2/Bax protein expression and inhibit endothelial cell apoptosis induced by high glucose through PI3k/Akt signal pathway,and thus may protect endothelial cells.

OBJECTIVESpinal muscular atrophy (SMA) is a common autosomal recessive disorder and represents one of the most common genetic causes of death in childhood. The last 10 years have seen major advances in the field of SMA, but no curative treatment is available so far. This study aimed to analyze the clinical characteristics of SMA, improve the clinical diagnosis of SMA, and explore the importance of gene diagnosis and prenatal diagnosis of SMA by gene deletion analysis.

METHODSTotally 83 cases with SMA including 55 males and 28 females were enrolled in this study. The age was between 1 day and 14 years (average 23.7 months). The clinical characteristics and changes of electromyography were assessed in all cases. The muscular biopsy was performed in 2 of 83 cases. The deletion of survival of motor neuron gene (SMN) was detected by PCR and restriction endonuclease spectrum analysis in 13 of 83 cases.

RESULTSThe 83 cases were subdivided into three clinical groups based on age of onset of symptom, age at death and achievement of certain motor milestone, 60 cases with type I, 19 cases with type II and 4 cases with type III. They were all characterized by symmetric muscle weakness (more proximal than distal) associated with atrophy, absence or marked decrease of deep tendon reflexes. Electromyographic studies showed a pattern of denervation with neither sensory involvement nor marked decrease of motor nerve conduction velocities in all cases. Muscle biopsy provided evidence of skeletal muscle denervation with groups of atrophy in 2 cases. The SMN detection revealed deletion of exon 7 and exon 8 in 11 of 13 cases, only lacking exon 7 in 1 of 13 cases and lacking exon 8 in 1 of 13 cases.

CONCLUSIONSMA is characterized by degeneration of lower motor neuron associated with muscle paralysis and atrophy. The definite diagnosis of SMA will rely on the typical clinical characteristics, changes of electromyogram and muscle biopsy and gene deletion analysis. Gene diagnosis of SMA can provide a basis for prenatal diagnosis which is of great importance in preventing SMA.

Adolescent ; Biopsy ; Child ; Child, Preschool ; Electromyography ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Muscle, Skeletal ; pathology ; Prenatal Diagnosis ; Spinal Muscular Atrophies of Childhood ; diagnosis ; genetics

Objective To observe the expressions of matrix metalloproteinase-9 mRNA and the change of cerebral edema after diffuse brain injury in rats and discuss their correlation.Methods Marmaruu's diffuse brain injury model of rat was made.Real-time quantitative RT-PCR,dry-wet meth- od,histological techniques and electron microscope were used to determine the expressions of MMP-9 containing water in brain tissue and inflammatory reaction and uhrastructural changes of blood capillary at different time phases after truama.Results The expressions of MMP-9 mRNA started to increase at 1 hour,peaked at 12 hours(P

Background Non-alcoholic fatty liver disease (NAFLD) is a complex disorder and has been closely linked to obesity.The fat mass and obesity-associated (FTO) gene is a newly discovered gene related to obesity,which enhances oxidative stress and tipogenesis in NAFLD.The forkhead transcription factor O1 (FoxO1) is another important gene involved in NAFLD,which causes lipid disorders when insulin resistance appears in the liver.However,the interactions between FTO and FoxO1 during the pathogenesis of NAFLD have not been fully elucidated.This study was designed to identify the relationship between these two factors that are involved in the development of NAFLD.Methods This study includes two parts referred to as animal and cell experiments.Twelve female SPF C57BL/6 mice were fed a high-fat diet to serve as an NAFLD animal model.Aspartate aminotransferase (AST),alanine aminotransferase (ALT),total triglyceride (TG),total cholesterol (TC),alkaline phosphatase (ALP),high-density lipoprotein (HDL),and low-density lipoprotein (LDL) were measured.Immunohistochemical analysis was used to detect the expression and histological localization of FTO,FoxO1,and adenosine monophosphate (AMP)-activated protein kinase (AMPK).The L02 cells were exposed to high fat for 24,48,or 72 hours.Oil red O staining was used to detect intracellular lipid droplets.Reverse transcription-polymerase chain reaction was used for analyzing the levels of FTO and FoxO1 mRNA.Results At the end of 10 weeks,ALP,ALT,AST,and LDL were significantly increased (P ＜0.01),while TC and TG were also significantly higher (P ＜0.05).In addition,HDL was significantly decreased (P ＜0.05).The FTO and FoxO1 proteins were weakly expressed in the control group,but both FTO and FoxO1 were expressed significantly higher (P ＜0.01) in the experimental group,and the expression of the two factors was significantly correlated.AMPK in the high-fat group showed a low level of correlation with FTO,but not with FoxO1.Oil Red O staining results showed that the cells cultured in 50％ fetal bovine serum for 24,48,or 72 hours exhibited steatosis.FTO and FoxO1 mRNA were increased in the high-fat group compared with the normal group (P ＜0.01).The expression levels of FTO and FoxO1 mRNA were the highest at 48 hours (P ＜0.05).Conclusions A high-fat diet leads to higher expression of FTO,phosphorylation of FoxO1,and decreased phosphorylation of AMPK.These results suggest that the interactions between FTO and FoxO1 are closely related to the pathogenesis of NAFLD.

OBJECTIVETo evaluate the effectiveness of locking plate external fixator in treating middle and distal tibial fractures.

METHODSFrom January 2010 to January 2013,18 patients suffered from middle and distal tibial fractures were treated by locking plate external fixator,including 11 males and 7 females, with an average age of 53.5 (ranged from 13 to 80) years old,the course of disease ranged from 2 h to 3 d. According to AO classification, 4 cases were type A,11 cases were type B and 3 were type C. Among them,6 patients were open fracture, including 2 cases with type I, 3 cases with type II and 1 case with type III, according to Gustilo classification), 12 patients were close fracture. Operation time, postoperative complications were observed, and Johner-Wruhs scoring were used to evaluate clinical outcomes.

RESULTSAll patients were followed up from 6 to 15 (meaned 11) months. Two cases occurred skin necrosis (1 case occurred bone exposure), 2 cases occurred delayed union (all were open fracture), and 1 case occurred nail infection. No screw loosening or broken occurred. According to Johner-Wruhs scoring, 10 cases obtained excellent result,6 cases good,and 2 cases fine.

CONCLUSIONLocking plate external fixator for the treatment of middle and distal tibial fractures, which has advantages of lessen damage, shorter operative time, less complications and rapid functional recovery, is one of good choice.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bone Plates ; utilization ; External Fixators ; utilization ; Female ; Fracture Fixation ; instrumentation ; methods ; Humans ; Male ; Middle Aged ; Tibial Fractures ; surgery ; Treatment Outcome ; Young Adult

Objective:To explore the efficacy and safety of laparoscopic colon cancer surgery for patients with end-stage renal disease .Methods:Thirty-one end-stage renal disease patients (continuous hemodialysis) underwent laparoscopic colon cancer surgery from Jan 2014 to May 2019 in Tianjin First Central Hospital compared to 35 colon cancer patients with normal renal function.Result:Compared with the control group, the operation time in laparoscopic group was longer[(187±20) min vs. (174±21) min, t=2.381, P=0.020], the intraoperative blood loss was more[90 (80-110) ml vs. 50(40-60) ml, Z=-6.580, P<0.001], the postoperative drainage volume was more[(417±89) ml vs.(208±67) ml, t=10.858, P<0.001], the postoperative hospitalization time was longer[(13.68±2.10) d vs.(9.09±1.65) d, t=9.918. P<0.001], and the total hospitalization costs were higher[9.2 (8.8-9.6) ten thousand yuan vs. 6.1 (5.8-6.5) ten thousand yuan, Z=-6.976, P<0.001]. There was no significant difference in overall morbidity between the two group (23% vs. 9%, P=0.170). Sixty-one patients (92%) were followed up for a median time of 6 months. One case of liver metastasis was found in each group. Conclusion:Laparoscopic colon cancer surgery can be a safe and effective procedure in patients with end-stage renal disease .

Objective:To explore the efficacy and safety of laparoscopy for acute appendicitis (AA)in kidney transplant(KT)recipients.Methods:From May 2016 to December 2022, retrospective review is conducted for 99 AA patients operated at Tianjin First Central Hospital.They are assigned into two groups of observation(kidney transplant recipients, 33 cases)and control(normal renal function patients, 66 cases). Laparoscopic perioperative data of white blood cell(WBC), neutrocyte percentage(NEUT%), C-reactive protein(CRP), procalcitonin(PCT), operative duration, intraoperative blood loss, postoperative drainage volume, postoperative hospitalization length, total hospitalization expense and morbidity of postoperative complications are compared.Results:After propensity score matching, no statistically significant inter-group difference existed in clinical profiles.WBC and NEUT% pre-operation are lower in observation group than those in control group[11.85(9.54~13.99)×10 9/L vs 13.74(12.42~14.66)×10 9/L, Z=-3.908, P<0.01; 85.00(73.65~89.60)% vs 88.20(83.85~90.20)%, Z=-2.522, P=0.012]. No significant inter-group difference existed in preoperative CRP/PCT(all P>0.05). No significant inter-group differences existed in WBC, NEUT, CRP or PCT at Day 3 post-operation(all P>0.05). No significant difference existed in creatinine level in observation group before and after operation( P>0.05). As compared with control group, operative duration was longer[86(74~99)vs 62(57~68)min, Z=-6.290, P=0.020], intraoperative blood loss greater[25(20~33)vs 15(15~20)ml, Z=-6.104, P<0.01], postoperative drainage volume larger[75(65~85)vs 35(25~36)ml, Z=-8.103, P<0.01], postoperative hospitalization time longer[7(6~9)d vs 5(4~5)d, Z=-7.064, P<0.01]and total hospitalization expense higher[(1.98±0.22)vs (1.73±0.22)ten thousand yuan, t=5.401, P<0.01]. No significant inter-group differences existed in time of postoperative passage of flatus( P=0.669). No significant inter-group difference existed in morbidity of postoperative complications( P=0.893). Conclusions:The efficacy of laparoscopic appendectomy in patients post-KT is basically comparable to that in counterparts with normal renal function.

The purpose of this study was to evaluate the safety of cryopreserved and thawed peripheral blood stem cell (PBSC) fractionated return infusions in children. 35 children patients with malignant tumors (13 acute leukaemias, 15 neuroblastomas and 7 malignant lymphomas) received fractionated return infusions of cryopreserved stem cells after undergoing high-dose chemotherapy without or with total body irradiation. The toxicities of 70 return infusions were evaluated. All patients were mobilized by chemotherapy plus recombination human granulocyte colony-stimulating factor (rhG-CSF), and then PBSCs were collected by a separator CS-3000 plus or COBE spectra-4. The grafts were cryopreserved in 10% dimethyl sulfoxide (DMSD) and stored in liquid nitrogen. There were totally 70 PBSC transfusions. The total volume of PBSCs transfused: 190 - 420 ml (265 +/- 73 ml or 13.7 +/- 4.2 ml/kg) with a mean of (4.43 +/- 1.91) x 10(8)/kg of PBSCs, and 0.94 +/- 0.18 g/kg of DMSO. The single dose: 90 - 300 ml (132 +/- 37 ml or 6.6 +/- 5.2 ml/kg) with a mean of 0.68 +/- 0.12 g/kg of DMSO. Symptoms occurring during the infusions were recorded. All patients were monitored for 24 hours after infusion. Pulse, blood pressure, body temperature, and respiratory rate were recorded every 15 minutes. At four hours before and 8 hours after infusion, urinalysis was performed. Serum potassium, sodium, creatinine, total bilirubin, aspartate amino transferase (AST), and alanine amino transferase (ALT) levels were examined within 24 hours before and after the first infusion. The results showed that the toxicities observed included hemoglobinuria in 54 return infusions (77.1%), headache in 28 (40.0%), nausea in 24 (34.3%), vomiting in 17 (24.3%), and abdominal pain in 8 (11.4%). Patients who received a graft > 200 ml tended to have a higher frequency of hemoglobinuria, headache, nausea, vomiting, or abdominal pain (P<0.01), and they disappeared quickly, too. Total bilirubin increased after the first return infusion (P<0.01), and there was a significant correlation between the volume of infusion and the degree of total bilirubin increase (r=0.8977, P<0.01). No renal failure or shock occurred. It is concluded that transient hemoglobinuria, headache, nausea, vomiting, and abdominal pain are common toxicities associated with PBSC autograft, and these toxicities are related with a single volume of PBSCs transfused. Total bilirubin increase is correlated with the volume of infusion. In a word, the toxicity is less frequent and lower severe in children with fractionated infusions of cryopreserved peripheral blood stem cell.
Acute Disease ; Adolescent ; Child ; Child, Preschool ; Cryopreservation ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Headache ; etiology ; Hematopoietic Stem Cell Mobilization ; methods ; Hemoglobinuria ; etiology ; Humans ; Leukemia ; therapy ; Lymphoma ; therapy ; Male ; Nausea ; etiology ; Neoplasms ; therapy ; Neuroblastoma ; therapy ; Peripheral Blood Stem Cell Transplantation ; adverse effects ; methods ; Recombinant Proteins

Cardiovascular disease (CVD) remains the leading cause of death worldwide. Therefore, exploring the mechanism of CVDs and critical regulatory factors is of great significance for promoting heart repair, reversing cardiac remodeling, and reducing adverse cardiovascular events. Recently, significant progress has been made in understanding the function of protein kinases and their interactions with other regulatory proteins in myocardial biology. Protein kinases are positioned as critical regulators at the intersection of multiple signals and coordinate nearly every aspect of myocardial responses, regulating contractility, metabolism, transcription, and cellular death. Equally, reconstructing the disrupted protein kinases regulatory network will help reverse pathological progress and stimulate cardiac repair. This review summarizes recent researches concerning the function of protein kinases in CVDs, discusses their promising clinical applications, and explores potential targets for future treatments.
Cardiovascular Diseases ; Heart ; Humans ; Myocardium ; Protein Kinases

Objective To investigate the molecular mechanisms of upregulated expression of cellular Fas-associated death domain-like interleukin-1 beta converting enzyme inhibitory protein(c-FLIP)by calreticulin(CRT)in patients with rheumatoid arthritis (RA). Methods The semi-quantitative analysis and localization of c-FLIP in RA and osteoarthritis (OA)synovium were detected by immunohistochemistry.The fibroblast-like synoviocytes(FLS)were isolated by enzymatic digestion of synovial tissue specimens obtained from RA and OA patients,and cultured as an in vitro experiment model.The expressions of c-FLIP in RA and OA synovial fibroblasts were detected by immunofluorescence and Western blot assay. Whether CRT influenced c-FLIP expression and its molecular mechanism were explored by Western blot assay. Results The high expression of c-FLIP was found in RA synovium, mainly in the lining and sublining areas of FLS and vascular endothelial cells detected by immunohistochemistry.Meanwhile,weak staining of c-FLIP was observed in OA synovium.The expression of c-FLIP was significantly higher in RA synovium than that of OA synovium(t=11.717,P<0.001).Results of immunofluorescence and Western blot assay showed that c-FLIP was mainly located in cytoplasm, and which was higher expressed in FLS of RA than that of OA. The increased c-FLIP expression and phosphorylation of NF-κB were detected after being co-incubated with exogenous CRT (0, 10, 50, 100 μg/L), in dose-dependent manner. The effect of CRT upregulating c-FLIP expression was blocked by NF-κB inhibitor BAY 11-7082.Conclusion CRT can increase c-FLIP expression at least partly through NF-κB pathway in RA,which may provide therapeutic target for the treatment of RA.