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Objective To evaluate whether patients aged over 70 years old is an independent predictor for prolonging the waiting time, and to know whether the population can be prioritized in the emergency grading system.Methods A retrospective analysis was conducted. All the clinical data of adult visitors to the emergency department of the Third Affiliated Hospital of Sun Yat-sen University from January 1st in 2016 to December 31st in 2017 were collected. All the patients received four emergency first aid triages and treated according to the emergency classification results. The relevant data of selected patients were retrieved from the hospital information system (HIS) including gender, age, triage level, triage department, outpatient service and initial diagnosis time. According to the conclusion of pre-phase analysis that the patients over 65 years old was not associated with waiting time, all subjects were divided into ≥ 70 years old group and < 70 years old group for further analysis. The waiting time was defined as the difference in registration time and the initial diagnosis time, and the patients were divided into two groups according to whether it exceeded the target waiting time (TWT). TWT was considered as follows according to each emergency triage level: level Ⅰ instantly, level Ⅱ10 minutes, level Ⅲ 30 minutes, level Ⅳ 60 minutes (subacute patients) or 2-4 hours (non-urgent patients). The multivariable Logistic regression analysis was conducted to determine whether the factor that aged over 70 years old was an independent predictor for prolonging waiting time over TWT.Results There were 272989 adult visits recorded totally, of which obstetrics and gynecology department visits, incomplete personnel information, invalid diagnosis (or no diagnosis), triage levelⅠ, abnormal waiting time (the waiting time exceeded 99％ of the average waiting time of all samples) records were excluded. Finally, 219299 emergency patients were enrolled, with 102218 male and 117081 female, which showed an average age of (35.86±15.81) years, 208082 patients (94.9％) in < 70 years old group, and 11217 (5.1％) in ≥ 70 years old group. Totally 191588 patients (87.4％) were diagnosed within the TWT, and the number was 86.5％ (9701/11217) in the ≥ 70 years old group. Multivariate Logistic regression analysis indicated that age under 70 years old was independently associated with an exceeded TWT [adjusted odds ratio (OR) = 1.106, 95％ confidence interval (95％CI) = 1.041-1.176,P = 0.001]. Meanwhile, visits during the peak period or low triage level was also a risk factor of exceeded TWT. According to the Logistic regression analysis of different departments and removing other factors, in the emergency internal medicine with the most elderly patients, the probability of exceeded TWT in ≥ 70 years old group was lessened (adjustedOR = 1.203, 95％CI = 1.121-1.291,P < 0.001).Conclusion In the emergency pre-grading system, the elderly aged (≥ 70 years old) was more likely to see a doctor in TWT than those under 70 years old, so they were easy to get priority treatment.
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Objective To study the expression of miR-146a in the serum of sepsis and the diagnostic value.Methods Serum miR-146a expression was detected in 98 patients diagnosed with sepsis,69 patients with severe sepsis and 30 healthy controls.ROC plots were used to evaluate the diagnostic value.Results The expression of miR-146a was decreased successively in healthy group,general sepsis group and severe sepsis group.In the diagnosis of sepsis,at the optimal expression cutoff miR-146a value of 0.805,the sensitivity was 0.967 and specificity 0.940,with an AUC of 0.983.In the diagnosis of sepsis severity degree,at the optimal expression cutoff miR-146a value of 0.530,the sensitivity was 0.796 and specificity 0.986,with an AUC of 0.943.Conclusion miR-146a can be a potential marker in diagnosis of sepsis and severity degree .
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AIM: In this study, CD147 antibody was used to carry out targeted modification of nanoparticles for protein kinase Cε (PKCε)-siRNA gene therapy to target lung cancer cells.The inhibitory effects of the nanoparticles on the proliferation and invasion of the lung cancer cells were observed.METHODS: The magnetic nanoparticles targeting CD147 protein were assembled as gene vector.The expression of CD147 in the lung cancer cells was observed under laser scanning confocal microscope.The cells were divided into CP group, CN group and LP group as the experimental groups. Targeted nanoparticles were used as CA group.Non-transfected cells were used as control group.The cell transfection was carried out with 250 ng plasmids/well in 6-well plate.The effect of nanocontrast agent on the cell endocytosis was observed under laser scanning confocal microscope.The mRNA expression of PKCε was detected by RT-qPCR.The protein expres-sion of Ki67, MMP3, PKCε, Wnt1 and GAPDH was determined by Western blot.The cell proliferation ability was detec-ted with colony formation assay.The cell invasion ability was detected by Transwell method.RESULTS: The expression of CD147 protein in the human lung cancer A549 cells was confirmed by immunofluorescence staining.The endocytosis of siRNA into the A549 cells in CP group was observed with the highest efficiency as compared with CN group and LP group. The relative mRNA expression of PKCε in the A549 cells of CP group, CN group, LP group and CA group were (9.76 ± 0.18)%, (98.51 ±0.32)%, (99.17 ±0.16)% and (99.68 ±0.11)%, respectively.The difference between CP group and control group was statistically significant (P <0.05).No significant difference among CN group, LP group and control group was observed.The protein expression of PKCε, Ki-67, MMP3 and Wnt1 in CP group was significantly reduced, and the protein expression levels among CN group, LP group and control group had no significant difference.The colony number in CP group was significantly smaller than that in control group (P <0.05).The effective colony numbers in CN group, LP group and CA group had no significant difference as compared with control group.The number of the invading cells in CP group was significantly less than that in control group (P <0.05).The numbers of the invading cells in CN group, LP group and CA group had no significant difference as compared with control group.CONCLUSION: Nanogene vector targe-ting CD147 can carry PKCε-siRNA to conduct gene therapy efficiently on the lung cancer cells to achieve effective inhibito-ry effects on the proliferation and invasion of the lung cancer cells.
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AIM:To investigate whether Notch1 changes stemness and chemotherapeutic sensitivity in human glioma U251 cells.METHODS: The lentiviral vectors, which expressed Notch1-shRNA or Notch1 intracellular domain ( NICD) , were transfected into U251 cells .Western blot and immunofluorescence staining were applied to monitor the va-lidity of the cells, down-regulation of Notch1 expression or over-expression of NICD.The proportion of CD133 +cells was analyzed by flow cytometry.The expression of nestin and GFAP was identified by immunofluorescence staining.The forma-tion rate of tumor cell spheres and the implanted tumor growth in SCID mice were observed.MTT assay was performed to e-valuate the chemotherapeutic sensitivity to VM-26 and BCNU of the cells with different treatments.RESULTS:Stemness was significantly enhanced in the cells over-expressing NICD.For example, the proportion of CD133 +cells was increased, the expression of nestin was up-regulated, the expression of GFAP was down-regulated, and the formation rate of tumor cell spheres and implanted tumor growth were increased.The chemotherapeutic sensitivity to VM-26 and BCNU of the cells was decreased.In the cells with Notch1 gene down-regulation by RNAi, the stemness was inhibited and chemotherapeutic sensi-tivity was increased.CONCLUSION:Notch1, which leads to the change of stemness and chemotherapeutic sensitivity in human glioma U251 cells, is likely to be a potential molecular target for treatment of glioma.
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<p><b>OBJECTIVE</b>To compare the safety of anastomosis and ostomy following 2-stage definitive colonic resection when severe colonic injuries treated in the setting of damage control surgery(DCS).</p><p><b>METHODS</b>Clinical data of 67 patients with severely traumatic colonic injuries undergoing DCS at the Third Affiliated Hospital of Sun Yat-sen University between 2005 and 2013 were analyzed retrospectively. Patients were divided into the anastomosis group undergoing colonic resection and anastomosis (n=40), and the ostomy group undergoing anastomosis with a protecting proximal ostomy (n=27). Postoperative complications were compared between these two groups. The risk factors of colonic anastomosis leakage were analyzed.</p><p><b>RESULTS</b>Demographics, injury severity, physiological imbalance on admission, transfusion during the first operative procedure were similar in the two groups (all P>0.05). Rates of anastomotic leakage, intra-abdominal abscess, enterocutaneous fistula, and would infection after definitive resection were not statistically different between the two groups (all P>0.05). Colonic anasomotic leakage rates were 15.0% (6/40) in anastomosis group and 11.1% (3/27) in ostomy group without significant difference (P>0.05). Left-sided colon injuries occurred in 7 out of 9 patients with anatomotic leakage, whose proportion was significantly higher than that in those without anastomotic leakage (7/9 vs. 24/58, 77.8% vs. 41.4%, P<0.05). A prolonged peritoneal closure was also observed in patients with anastomotic leakage (median, 10 days vs. 2 days, P<0.05).</p><p><b>CONCLUSIONS</b>A strategy of diverting ostomy is not the first choice for patients suffering from severe colonic injuries in the setting of DCS. Peritoneal closure at early stage may decrease the risk of colonic anastomotic leakage.</p>
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Humans , Abdominal Abscess , Abdominal Injuries , Anastomosis, Surgical , Anastomotic Leak , Colonic Diseases , General Surgery , Postoperative Complications , Retrospective Studies , Risk FactorsABSTRACT
AIM:To investigate the inhibitory effect of small interfering RNA ( siRNA) on the expression of protein kinase Cε( PKCε) in human hepatoma SK-Hep-1 cells, and the biological behaviors of the transduced cells , inclu-ding proliferation and invasion , were investigated.METHODS:The cultured SK-Hep-1 cells were divided into 3 groups, including PKCε-siRNA group , negative control ( NC)-siRNA group and control group .MTT assay was used to analyze the proliferation of the SK-Hep-1 cells in the respective groups , while invasion potency was determined by Transwell assay .The protein levels of functional biomarkers such as Ki 67 and matrix metalloproteinase 9 ( MMP-9 ) were measured by Western blotting .The Luciferase reporter gene assay was used to explore the activity of the NF-κB pathway .RESULTS:PKCεex-pression in SK-Hep-1 cells transfected with PKCε-siRNA was significantly down-regulated at both mRNA and protein levels compared with that in the normal SK-Hep-1 cells (P<0.01), with the decreases in the protein levels of Ki67 and MMP-9. The invasion and proliferation of SK-Hep-1 cells were obviously inhibited in PKCε-siRNA group compared with control group (P<0.01).Furthermore, the transcriptional activity of NF-κB was down-regulated when PKCε was effectively in-hibited by PKCε-siRNA (P<0.01).CONCLUSION:Down-regulation of PKCεinhibits the proliferation and invasion of hepatic carcinoma cells , which might be mediated via the NF-κB signaling pathway .
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Objective To analyse the frequeney and pattern of lymph node metastasis in bilateral papillary thyroid microcarcinoma (PTMC), and establish the optimal surgical strategy for patients. Methods From March 2006 to August 2008, 58 bilateral PTMC patients received surgical treatment and the tumour characteristics, the frequency and pattern of lymph node metastasis and surgical management of these patients were retrospectively analysed. Results Forty-four patients received total thyroideetomy and 14 patients received near-totsl thyroideetomy, 47 patients received central compartment (level VI ) dissection and cervical level Ⅱ,Ⅲ, IV node exploration by internal jugular vein exposure,10 patients received level Ⅵdissection and unilateral cervical dissection and 1 patient received bilateral cervical dissection. The mean tumor diameter was (6.28 + 2.23) mm and 26 patients (44.8%) had node involvement, 88.5%(23/26) pa-tients had only level Ⅵ node involvement. Only 1 patient had node involvement in the jugular chain without level Ⅵ node involvement, 2 patients with level Ⅵ node involvement were associated with another cervical compartment nodes involvement. Conclusions Bilateral PTMC has high incidence of lymph node metasta-sis. The cervical level Ⅵ is the most common site of node involvement for bilateral PTMC and the surgical strategy for bilateral PTMC should include the cervical level Ⅵ dissection routinely.
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<p><b>OBJECTIVE</b>To understand the pathogenic effect and the correlation between the genotype and phenotype of the 4 novel missense mutations (G247S, E280G, P362T and A434D) of phenylalanine hydroxylase gene (PAH).</p><p><b>METHODS</b>(1) The enzyme activity of the 4 mutants was assessed by using transient protein expression in mammalian cells. (2) The PAH amino acid sequences among different animal species were alignmented. (3) The effects of the 4 missense mutations on the protein structure were analyzed. (4) The clinical phenotype of the patients with PKU were analyzed, according to their blood Phe levels prior to treatment and the Phe tolerance.</p><p><b>RESULTS</b>(1) The residual enzyme activity expressed in vitro of G247S, E280G, P362T and A434D were 3.1%, 0.4%, 8.2% and 21.7% of the wild-type PAH respectively; (2)Gly247, Glu280 and Pro362 were among the highly conserved amino acids, while Ala434 was only moderately conserved; (3) As revealed by 3D structural analysis, G247S and E280G, being located at the active center of the enzyme, interfered with the binding of PAH to BH4 and ferrousion respectively, while P362T and A434D affected the formation and stability of the dimer and the tetramer of PAH; (4) As shown by clinical phenotypic analysis, classical PKU were observed in patients carrying G247S and E280G, moderate PKU were observed in patients carrying A434D, whereas both classical and moderate PKU were observed in patients carrying P362T.</p><p><b>CONCLUSION</b>(1) The E280G, G247S, P362T and A434D are all disease-causing mutations, with those located at the center of the enzyme displaying the most marked pathogenic effect; (2)The results of the structural analysis of the 3D molecule are consistent with the activity assessment of the enzyme expressed in vitro; (3) The consistency is observed between the genotype, the enzymatic activity expressed in vitro and the clinical phenotype.</p>
Subject(s)
Animals , Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Male , Amino Acid Sequence , Gene Expression , Genotype , Models, Molecular , Molecular Sequence Data , Mutant Proteins , Chemistry , Genetics , Metabolism , Mutation, Missense , Phenotype , Phenylalanine Hydroxylase , Chemistry , Genetics , Metabolism , Phenylketonurias , Genetics , Protein Conformation , Sequence Alignment , Structure-Activity RelationshipABSTRACT
OBJECTIVE To compare two different observed data from Nov 2003 to Apr 2004 and from Jan 2005 to Dec 2005 on hospital-acquired infection in our hospital and put forward the counterned measures.METHODS The bacteria samples from clinic were cultured.RESULTS The Pseudomonas aerugonosa,Acinetobacter baumannii and Escherichia coli were common strains of bacteria according to the two years data in our hospital.They were 30.35%,18.41% and 8.46% vs 19.74%,17.11% and 14.04%,respectively.The P.aerugonosa infective constituent ratio was decreased(30.35% vs 19.74%,P
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Objective:To observe the sensitivities of breast cancer cells and its implanted tumors to chemotherapeutic drug epirubicin after down-regulation of HER-2 expression by RNA interference(RNAi).Methods:HER-2siRNApU6 vector containing HER-2 RNAi was constructed and was used to transfect HER-2 positive breast cancer cell line SKBR-3.The expression of HER-2 mRNA and protein were analyzed by RT-PCR and Western blotting,respectively.Transfected SKBR-3 cells were treated with different concentrations of epirubicin;the growth of SKBR-3 cells and IC50 of epirubicin were observed by MTT.SKBR-3 cells were injected into nude mouse to establish breast cancer model;the sensitivity of mouse model to epirubicin was observed after HER-2shRNApU6 treatment.Results:Expression of HER-2 mRNA and HER-2 protein were down-regulated in SKBR-3 cells after transfection with HER-2shRNApU6.Furthermore,the proliferation of SKBR-3 cells transfected with HER-2shRNApU6 was significantly decreased compared with mock tansfected group(P