Objective:To investigate the value of bedside transthoracic echocardiography(TTE) in volume reactivity assessment of children with septic shock.Methods:A total of 41 children aged from 1 to 5 years with septic shock requiring mechanical ventilation admitted to PICU from January 2017 to June 2020 were prospectively included.Under the condition of complete mechanical ventilation, full sedation and analgesia, and no spontaneous breathing(tidal volume 8 to 10 mL/kg), volume expansion was given to children.Hemodynamic indexs such as cardiac index(CI), stroke volume index(SVI) and stroke volume variability(SVV) were measured before and after volume expansion by noninvasive cardiac output monitoring(NICOM) and TTE.Moreover, aortic flow velocity time integral variable degrees(ΔVTI), inferior vena cava variability(ΔIVC) and inferior vena cava dilation index(dIVC) were also measured by TTE.Patients were considered to be responsive to volume expansion if SVI NICOMincreased≥15%.Based on the responsiveness of volume expansion, all the patients were divided into response group and non-response group.The value of SVV TTE, ΔVTI, ΔIVC, dIVC, ΔCVP and SVV NICOMin predicting volume responsiveness were analysed. Results:(1) There were 23 cases in response group and 18 cases in non-response group.Before volume expansion, there were no statistically significant differences in general hemodynamic indexes HR, MAP, CVP, EF, CI NICOM, and CI TTEbetween two groups( P>0.05). (2) In response group, HR, MAP, CI, SVI and CVP were all improved after volume expansion( P<0.001). In non-response group, only CVP was significantly increased after volume expansion, while other indexes were not improved( P>0.05). (3)Before the volume expansion, SVV TTE, ΔVTI, ΔIVC, and dIVC in response group were higher than those in non-response group( P<0.001). After volume expansion, these indicators were significantly reduced in response group.In non-response group, only ΔIVC significantly reduced after volume expansion.(4) The receiver-operating characteristic curve analysis showed that the area under the curve of SVV TTEand ΔVTI was 0.971, with 12.04% as the threshold, the sensitivity was 0.957 and the specificity was 0.944. The area under the curve of ΔIVC was 0.981, with 25.98% as the threshold, the sensitivity was 0.870 and the specificity was 1.000.The area under the curve of dIVC was 0.980, with 29.86% as the threshold, the sensitivity was 0.870 and the specificity was 1.000. The area under the curve of ΔCVP was 0.778, with 2.5 cmH 2O(1 cmH 2O=0.098 kPa) as the threshold, the sensitivity was 0.913 and the specificity was 0.556. The area under the curve of SVV NICOMwas 0.874, with 12.50% as the threshold, the sensitivity was 0.869 and the specificity was 0.778. Conclusion:The dynamic indexes SVV, ΔVTI, ΔIVC and dIVC monitored by TTE have good accuracy in evaluating children′s volume responsiveness, among which the accuracy of ΔIVC and dIVC is relatively the highest; the value of ΔCVP in predicting volume responsiveness is limited.

Objective To analyze the possible reasons of failed surgical treatment of acetabular fractures. Methods Various methods were used for positive patient identification, including according to Matta's X-ray assessment and Merle d'Aubigne & Postel hip function score of clinical standards for classification of acetabular fracture reduction surgery were not satisfied or not carried out a reduction and fixation,the clinical evaluation of hip joint as a "bad", occurrence of femoral head subluxation or dislocation, femoral head necrosis and other serious complications. From February 2000 to February 2008, 22 patients including 14 males and 8 females with an average age of 38.6 years (range, 18-72 years) were considered as failed cases. Results 45.5% of these cases were posterior wall fractures which were not given any fixation, 27.3% of them were posterior column fractures which were not fixed, 13.6% of them whose reduction and fixation of anterior wall fractures were not satisfied, and 9.1% of them were anterior column fractures which needed fixation. One case should take open reduction and iternal fixation instead of THA. The rate of misdiagnosis and mistaken diagnosis were 90% if only X-ray evaluation was made and this rate decreased to 8.3% if computed tomography was taken. The rate of wrong selection of operative approach was 100% in 10 cases of misdiagnosis, and which was 58.3% in 12 cases of correct diagnosis. In the 5 patients with correct diagnosis and selection of operative approach, the reasons of failed surgical treatment were due to imperfect surgical skills in 3 cases, and inappropriate fixation patterns in 2 cases. Conclusion The causes of the failure of surgical treatment for acetabular fracture might include preoperative missed diagnosis and misdiagnose, inappropriate approach, and an unreasonable internal fixation with unskillful technique.

OBJECTIVE: Through preoperative temporary balloon occlusion of internal carotid artery and monitoring of carotid artery stump pressure variation, in order to further predict the risk of carotid artery ligation and resection, evaluation operative risk and provides the reference for the choice of surgical approach. METHOD: Continuous monitoring and recording the carotid artery stump return pressure,before clamping and in the process of blocking, close observation the patients mental state and the nervous systemof all kinds of signs, in the process of blocking, to understand the dynamic change of stump artery pressure return in patients and whether can the smooth passage of carotid artery balloon occlusion test. RESULT: Of the 19 patients, 4 cases were positive, 15 negative cases, Blocking immediate the positive patients and negative patients with stump pressure drop was (57. 35 ± 1. 89) % and (38. 99 ± 12. 23) %, with statistical significance between the two, in the process of blocking, the mean stump pressure of the positive patients and the negative patients was (37. 29 ± 3. 15) mmHg and (61. 36 ± 14. 69) mmHg, with statistical significance between the two. CONCLUSION Approximately 21. 05% of patients can not tolerate carotid artery balloon occlusion test, theory for carotid artery reconstruction operation. After blocking the stump pressure is less than 40. 44 mmHg, the theory for reconstruction of the internal carotid artery operation. Blocking instant artery stump pressure dropped more than 55. 46%, in theory the need for internal carotid artery reconstruction.
Balloon Occlusion ; Blood Pressure ; Carotid Artery, Internal ; surgery ; Feasibility Studies ; Humans ; Ligation ; Preoperative Care ; Risk Assessment ; Vascular Surgical Procedures

OBJECTIVETo investigate the effects of sphingosine kinase 1 (SphK1) on the proliferation, migration and invasion of human colon cancer LoVo cells, and to explore the related mechanisms.

METHODSHuman colon cancer LoVo cells were divided into three groups: phorbol 12-myristate 13-acetate (PMA) was used to induce the activation of SphK1 in the PMA group, N,N-dimethylsphingosine (DMS) used to suppress the activity of SphK1 in DMS group, and the cells treated with equal amount of 0.9 % NaCl instead of drugs served as the control group. The activity of SphK1 was assayed by autoradiography, the cell proliferation was assessed by MTT assay, cell migration and invasion were examined by Boyden chamber assay, concentrations of sICAM-1 and sVCAM-1 were assayed by ELISA, and RT-PCR and Western blot were used to evaluate the mRNA and protein expression in the cells.

RESULTSThe activity of SphK1 was efficiently induced by PMA and significantly suppressed by DMS. PMA induced cell proliferation in a time- and dose-dependent manner. On the contrast, DMS suppressed cell proliferation in a time- and dose-dependent manner. After treating with PMA, the number of migrating and invasing cells were increased to 143.36 ± 8.73 and 118.46 ± 6.25, significantly higher than those of the control group (75.48 ± 6.12 and 64.19 ± 5.36). After treating with DMS, the number of migrating and invasing cells were decreased to 38.57 ± 3.24 and 32.48 ± 4.27, significantly lower than those of the control group (P < 0.01). The relative expression levels of FAK, ICAM-1 and VCAM-1 mRNA in the PMA group were 0.82 ± 0.06, 0.74 ± 0.05 and 0.89 ± 0.09, and those in the DMS group were 0.23 ± 0.02, 0.26 ± 0.03 and 0.37 ± 0.04, with significant differences between the PMA, DMS and control groups (P < 0.01). Compared with the control group, the relative expression levels of FAK and p-FAK proteins in the PMA group (0.52 ± 0.06 and 0.51 ± 0.06) were significantly elevated, and those of the DMS group (0.20 ± 0.03 and 0.09 ± 0.02) were significantly decreased. In addition, the concentrations of sICAM-1 and sVCAM-1 were significantly elevated with the activation of SphK1. On the contrary, those of the DMS group were significantly reduced with the suppression of SphK1 (Both P < 0.01).

CONCLUSIONSSphK1 may enhance the proliferation, migration and invasion of colon cancer LoVo cells through activating FAK pathway and up-regulating the expression of ICAM-1 and VCAM-1.

Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Colonic Neoplasms ; enzymology ; metabolism ; pathology ; Dose-Response Relationship, Drug ; Enzyme Inhibitors ; pharmacology ; Focal Adhesion Kinase 1 ; genetics ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; metabolism ; Neoplasm Invasiveness ; Phosphorylation ; drug effects ; Phosphotransferases (Alcohol Group Acceptor) ; metabolism ; RNA, Messenger ; metabolism ; Signal Transduction ; Sphingosine ; analogs & derivatives ; pharmacology ; Tetradecanoylphorbol Acetate ; pharmacology ; Vascular Cell Adhesion Molecule-1 ; genetics ; metabolism

To investigate the influence of HIF-1alpha overexpression on the differentiation of endothelial progenitor cells (EPCs) ex vivo, EPCs were isolated from human peripheral blood by density gradient centrifugation, overexpressed HIF-1alpha was transfected to EPCs by electroporation; HIF1alpha, HIF1beta, vascular endothelial growth factor (VEGF) mRNA level were measured with RT-PCR; HIF-1alpha protein was detected with immunohistochemistry in a time course. CD31(+) cells were measured with flow cytometry. Cell morphology was observed after transfection. The results showed that the transfection efficiency of HIF-1alpha to EPCs was about 20%. HIF-1alpha and its controlled target gene VEGF were markedly induced by HIF-1alpha vector (P < 0.05). HIF1beta had its same level as it before interference (P > 0.05). HIF-1alpha protein was induced by HIF-1alpha transfection after 12 hours but was undetectable at 24 hours. After 7 - 14 days cultured in 21% oxygen pressure, fluorescence-trace experiments revealed that CD31 + EPCs/EC could be generated more efficiently from overexpressed HIF-1alpha than that from pEGFP transfected group (P > 0.05). EPC morphology was observed by light microscopy. HIF-1alpha-transfected cells under normoxia sprouted more rapidly from the EPC colonies than the untransfected cells or cells transfected with an GFP vector, which essentially maintained the original colony formation. HIF-1alpha transfected cells took on an array-like arrangement rather than random dispersal, suggesting that they were in an advanced state of differentiation. It is concluded that the utility of overexpression of HIF-1alpha can induce target genes which have influence on cell differentiation. HIF-1alpha transfection was found to give a prospected way to do the insight research on ischemic treatment in vivo.
Cell Differentiation ; drug effects ; Endothelial Cells ; cytology ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; biosynthesis ; genetics ; Plasmids ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Stem Cells ; cytology ; Transfection ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

Multiplex reverse transcription-polymerase chain reaction (mRT-PCR) is currently available in virus detection and defined as the simultaneous amplification of two or more DNA/RNA targets in a single reaction vessel. In this study, we attempted to modify the conventional mRT-PCR technique on a basis of GenomeLab Genetic Analysis System (GeXP). Initially, we optimized the analytical validation of the GeXP analyzer and its design of workflow and simultaneously detected eight arboviruses that related to epidemic encephalitis by verifying the specificity of mRT-PCR with Japanese encephalitis virus(JEV) cell cultures and positive strains identified previously and determining the sensitivity with in vitro-transcribed RNA of serial dilutions. The GeXP system after optimization could amplify the specific fragments related to the viruses and exposed specifically a total of 13 target genes out of eight types of arboviruses at the level of 10(2) copies/microL, and the findings suggest that the novel protocol we developed can be high-throughput and highly specific and sensitive as well as quickness in screening of the encephalitis viruses, and is promising in detection of encephalitis-associated viruses for molecular epidemiological studies.
Arboviruses ; genetics ; isolation & purification ; Encephalitis Virus, Japanese ; genetics ; Encephalitis Viruses ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

OBJECTIVEAutophagy is involved in several neurodegenerative diseases and recently its role in acute brain injury has won increasing interest. Spinal cord injuries (SCIs) often lead to permanent neurological deficit. Therefore, in this study, we examined the pro?les of autophagy-linked proteins (MAP-LC3) after SCI to investigate whether the expression of autophagy contributes to neurological deficit after SCI.

METHODSAdult female Sprague-Dawley rats were used and randomly divided into control and SCI groups. All the rates received laminectomy at T8-T10 level. Those in the SCI group received additional exposure of the dorsal surface of the spinal cord, followed by a weight- drop injury. Thereafter we investigated the expression levels of MAP-LC3, beclin-1, Cathepsin D and the beclin-1-binding protein bcl-2 by western blot analysis at 12 h, 24 h, 3 d, 7 d, 21 d and 28 d. One-way ANOVA with Tukey post hoc test was used to compare data between groups.

RESULTSWe observed significant increase in the level of LC3 (LC3-II/LC3-I) at 3 d and 7 d after SCI when compared with the sham group. While the level of beclin-1 and ratio of beclin-1/bcl-2 was found to have increased from 12 h to 24 h after injury. Cathepsin D expression was also elevated at 7 d (P<0.01).

CONCLUSIONBased on the above mentioned data, we proposed that autophagy plays a role in the manifestation of cell injury following SCI.

Adaptor Proteins, Signal Transducing ; metabolism ; Animals ; Apoptosis Regulatory Proteins ; metabolism ; Autophagy ; physiology ; Beclin-1 ; Blotting, Western ; Cathepsin D ; metabolism ; Disease Models, Animal ; Female ; Laminectomy ; Microtubule-Associated Proteins ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; metabolism

This study was aimed to investigate the effect of rhIL-6 and rhEPO on hepcidin mRNA expression in HepG2 cells and human primary hepatocytes, and mechanism of rhEPO in treatment of anemia of chronic disease (ACD). The HepG2 cells and human primary hepatocytes were cultured with medium containing different concentrations of rhIL-6 and rhEPO for a certain time, then mRNA was isolated and its RT-PCR was performed, the bands were photographed and analyzed by UVI band, the hepcidin and G3PDH mRNA ratio were semi-quantitatively analyzed. The expression levels of hepcidin in GepG2 cells and human primary hepatocytes at different conditions were compared. The results showed that the hepcidin mRNA expression in HepG2 cells and human primary hepatocytes could be enhanced by rhIL-6, the rhEPO could inhibit rhIL6-induced hepcidin mRAN expression. The rhEPO alone basically did not influence hepcidin mRNA expression in HepG2 cells. It is concluded that Hepcidin mRNA expression in HepG2 cells and human primary hepatocytes can be elevated by rhIL-6 with concentration- and time-dependent manner in certain range. rhEPO can inhibit this effect of rhIL-6.
Antimicrobial Cationic Peptides ; genetics ; metabolism ; Erythropoietin ; pharmacology ; Hep G2 Cells ; Hepatocytes ; drug effects ; metabolism ; Hepcidins ; Humans ; Interleukin-6 ; pharmacology ; RNA, Messenger ; genetics ; Recombinant Proteins ; pharmacology

AIMTo study the chemical constituents of Ziziphus jujuba Mill var. Spinosa (Bunge) Hu ex. H. F. Chou.

METHODSTo separate the constituents by using various kinds of chromatography and identify their structures on the basis of spectral analysis.

RESULTSFive compounds were isolated and their structures were established as jujuboside D (1), jujuboside A (2), 5,7,4'-trihydroxyflavonol-3-O-beta-D-rhamnopyranosyl-(1-->6)-beta-D-glucopyranoside (3), 6'''-coumaroylspinosin (4) and phenylalanine (5).

CONCLUSIONCompound 1 is a new compound named jujuboside D, 4 is reported as rotamer for the first time, 3 and 5 isolated from this plant for the first time.

Molecular Conformation ; Molecular Structure ; Phenylalanine ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Saponins ; chemistry ; isolation & purification ; Seeds ; chemistry ; Ziziphus ; chemistry

OBJECTIVETo investigate the effect of sphingosine kinase 1 (SphK1) on the proliferation, apoptosis, migration and invasion of colon cancer TH-29 cells and to explore its molecular mechanisms.

METHODSPhorbol 12-myristate 13-acetate (PMA) was used to induce the activity of SphK1 and N, N-dimethylsphingosine (DMS) was used to suppress the activity of SphK1. Cell prolieration and apoptosis were detected by MTT assay and flow cytometry, respectively. The migration and invasion capabilities of the cells were assessed in Transwell chambers. The activity of SphK1 was assayed by autoradiography. Western blot was used to evaluate the protein expression of SphK1, p38, phosphorylated p38 (p-p38) and SAPK/JNK.

RESULTSPMA and DMS were able to induce and suppress the activity and protein expression of SphK1 in a time-dependent manner, respectively. PMA enhanced and DMS suppressed the cell viability in a time- and dose-dependent manner. Being treated with 100 nmol/L PMA or 50 µmol/L DMS for 0, 6, 12, 24 h, the cell apoptosis rates of PMA group were (9.35 ± 0.84)%, (7.61 ± 0.48)%, (5.53 ± 0.76)% and (0.56 ± 0.33)%, contrastly, that of DMS group were (9.18 ± 0.94)%, (12.06 ± 1.41)%, (19.80 ± 2.36)% and (31.85 ± 3.60)%, respectively. Compared with the control group, the cell migration and invasion capabilities of the PMA group were significantly enhanced, and that of the DMS group were significantly suppressed. The migration cell number of control, PMA and DMS groups were 68.75 ± 6.15, 109.33 ± 11.63 and 10.83 ± 2.48, the invasion cell number of control, PMA and DMS groups were 55.42 ± 4.50, 90.58 ± 7.06 and 9.58 ± 2.39, respectively. With the elevating activity and expression of SphK1, the protein expressions of p38, p-p38 and SAPK/JNK were strikingly suppressed. On the contrary, after treating with DMS the protein expressions of p38, p-p38 and SAPK/JNK were enhanced.

CONCLUSIONSSphK1 potently enhances the prolieration, migration and invasion of colon cancer HT-29 cells, meanwhile suppresses the cell apoptosis. The suppressing of the p38 and SAPK/JNK signalling pathways may be one of its molecular mechanisms.

Apoptosis ; drug effects ; Carcinogens ; administration & dosage ; pharmacology ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Enzyme Inhibitors ; administration & dosage ; pharmacology ; HT29 Cells ; Humans ; MAP Kinase Kinase 4 ; metabolism ; Neoplasm Invasiveness ; Phosphorylation ; Phosphotransferases (Alcohol Group Acceptor) ; metabolism ; physiology ; Sphingosine ; administration & dosage ; analogs & derivatives ; pharmacology ; Tetradecanoylphorbol Acetate ; administration & dosage ; analogs & derivatives ; pharmacology ; Time Factors ; p38 Mitogen-Activated Protein Kinases ; metabolism