RESUMEN
Objective @#To elucidate the effect of phosphorylation modification at the threonine 592 (Thr592) site on the inhibition of gastric cancer proliferation by sterile alpha motifs and HD structural domain－containing protein 1 (SAMHD1) and the potential mechanism of action．@*Methods@# Post-translational modifications (PTMs) of SAMHD1 protein in gastric cancer tissues and cell lines in the database were analyzed，and immunohistochemical stai- ning was performed to detect SAMHD1 Thr592 phosphorylation in paired tissues of gastric cancer patients．In gastric cancer cells，SAMHD1 Thr592 variants were constructed and transiently transfected，and cell proliferation was detected using the cell counting kit 8 ( CCK-8 ) method． The phosphorylation of the cyclin-dependent kinases ( CDK) 2 protein threonine 160 (Thr160) site was inhibited by the addition of different concentrations of the CDK6 inhibitor，Palbociclib，which reduced the level of SAMHD1 protein Thr592 phosphorylation．Three online databases were used to analyze the SAMHD1 reciprocal proteins and take the intersection to derive the Nik-related kinase (NRK) protein．Immunoprecipitation ( Co-IP) ，mass spectrometry and Western blot were used to verify the interactions between SAMHD1 and NRK proteins and detect the effect of NRK on the phosphorylation of the SAMHD1 Thr592 site. @*Results @#Compared with PTMs such as ubiquitination，the highest level of phosphorylation modification of SAMHD1 was observed in tumors，and the difference was statistically significant (P＜0. 01) ．Immunohistochemical experiments showed that phosphorylated SAMHD1 (Thr592) was expressed higher in gastric adenocarcinoma than that in normal mucosal tissue adjacent to the cancer，and the difference was statistically significant (P < 0. 01) ．Western blot assay showed that SAMHD1 protein expression was elevated in MKN-45 cells in the overexpression wild type and mutant groups ，and phosphorylated SAMHD1 levels were also elevated in the wild type， T592E and HD / AA groups． CCK-8 assay showed that both SAMHD1 wild type and T592A could inhibit gastric cancer cell proliferation，while T592E and HD / AA had no effect on gastric cancer proliferation． On the basis of overexpression of SAMHD1，CCK-8 suggested that cell proliferation was inhibited after adding different concentrations of Palbociclib treatment，and Western blot assay suggested that the phosphorylation level was also reduced. NRK protein was obtained by Co-IP and mass spectrometry identification to screen the SAMHD1 reciprocal protein profile and database intersection，and NRK was found to interact with SAMHD1 protein and promote phosphorylation at SAMHD1 Thr592 site by Co-IP and Western blot assay．@*Conclusion @#Phosphorylation of the Thr592 site contributes to the loss of SAMHD1 's ability to inhibit gastric cancer cell proliferation，which is reversed by Palbociclib．NRK interacts with SAMHD1 protein，promoting phosphorylation of the SAMHD1 Thr592 site.
RESUMEN
Objective To investigate clinical characteristics and prognostic factors for hospitalized patients aged 95 years and older with community-acquired pneumonia(CAP).Methods A retrospective study was conducted on CAP patients aged ≥95 years hospitalized in Jiangsu Province Hospital or Jiangsu Province Hospital of TCM between January 2014 and January 2018.Clinical characteristics were collected.The shock index (SI),Charlson comorbidity index (CCI),neutrophil/lymphocyte rate (NLR)and CURB-65 score were calculated.The predictive value of the related factors was evaluated by using the Logistic regression model and the subjects' receiver operating characteristic curve (ROC).Results A total of 205 cases were enrolled in this study.The hospital mortality rate was 36.1 ％ (73/205).The heart rate,white blood cell count,mononuclear cell count,neutrophil count (N),NLR,fasting blood glucose,blood urea nitrogen(BUN) and D-dimer results revealed significant differences between the death group and the survival group(all P＜0.05).Scores of CCI and CURB-65 were higher in the death group than in the survival group[(2.90 ±1.88) vs.(1.91 ± 1.81),(2.34 ±0.69)vs.(1.76±0.69),both P＜0.05].Multivariate Logistic analysis showed that heart rate≥84.73 beat per minute(OR =2.452,95％CI:1.054～5.702),NLR≥11.43(OR =5.499,95％CI:2.132～14.180),BUN≥12.95 mmol/L(OR =2.546,95％CI:1.025～6.319),CCI≥3(OR =4.453,95％CI:1.191～10.113),CURB-65≥2 scores(OR =3.888,95％CI..1.288～11.735)and respiratory failure (OR =2.875,95 ％CI:1.204～6.806)were risk factors for death in hospitalized elderly aged 95 years and older with CAP.ROC analysis showed that CCI combined with the CURB-65 score or CCI combined with the CURB-65 score plus NLR could better predict the in-hospital mortality of elderly patients than CCI or the CURB-65 score used alone.Conclusions Hospitalized patients aged≥95 with CAP are prone to multiple complications and high mortality rates.Combined application of multiple evaluation systems and indicators can improve the prognosis of outcomes for patients in this age group.
RESUMEN
Objective To investigate the health status of centenarian hospitalized patients and analyze the risk factors for in-hospital death in Nanjing district.Methods All centenarians hospitalized patients who were discharged from wards of 10 upper first-class general hospitals in Nanjing district during the past five years were retrieved from their hospital information systems.Then,a retrospective study was performed on centenarians' data of general information,laboratory test results,Charlson comorbidity index (CCI),neutrophil to lymphocyte ratio (NLR) and shock index(SI),etc.were calculated and collected.Relevant risk factors for in-hospital death were analyzed by multivariate logistic regression analysis.Results A total of 156 patients aged 100 years and over,with an average age of (101.0±2.1)years,were enrolled during the past 5 years.The top 3 admitting diagnosis for the patients were pulmonary infection(30.1％,47/156 cases),coronary heart disease(10.9％,17/156 cases)and cerebrovascular disease(7.1％,11/156 cases).Fifty patients died during hospitalization,with a mortality of 32.1％ (50/156).Pneumonia was the most common admitting diagnosis(40.0％,20/50 case).Among causes of death,the combined admitting diagnosis with dementia,chronic renal insufficiency,one or more basic disease were significantly associated with death.There were statistically significant differences between bad vs.good vs.indifferent prognosis in heart rate,shock index,leukocyte count,neutrophil count,NLR,hemoglobin,albumin,albumin/globulin,fasting blood glucose,blood urea nitrogen,serum creatinine,C-reactive protein(CRP)and CCI levels.Multivariate logistic regression analysis suggested that NLR≥13.18,fasting blood glucose ≥7.56 mmol/L,blood urea nitrogen ≥20.74 mmol/L,CRP≥65 mg/L and CCI≥3 might be predictors for in-hospital death in the cohort(OR =48.91、3.43、1.22、6.55、1.55,all P＜0.05).Conclusions Pulmonary infection is the most common reason for admission and the cause of death in centenarian inpatients.Comorbidities increase the risk of death.To lower in-hospital mortality,CCI and other assessment indicators should be used to strengthen the comprehensive assessment and chronic disease management of hospitalized centenarians.Infectious diseases should be prevented beforehand.
RESUMEN
Objective To summarize the method of forearm skin flap with dorsal interosseous artery placed by turning the proximal end of the flap to the distal side of the wound to repair skin and soft tissue defect of palm and thumb of hand,and put forward the key points of operation.Methods From 2008 to 2013,the forearm skin flap with interosseous dorsal artery to repair skin and soft tissue defect of palm and the part between thumb and index finger in 20 cases.Dorsal interosseous artery in the forearm is passing through between superficial and deep layers of extensor muscles and sends out 5-13 cutaneous branches to dorsal antebrachium skin.The large skin flap has good blood supply with high survival rate.Results All flaps survived,and good results were achieved in all 20 cases.Conclusion The flap preserved two main arteries from upper limb with good blood supply,and the amount of blood supply of the flap depended on the size of flap,and the wound of donor site can be directly sutured up or covered with free skin graft,thus minimizing the alteration of appearance and function of donor area.
RESUMEN
AIM:To investigate the effect of caspase-8 small hairpin RNA ( shRNA) on attenuating apoptosis of human mesenchymal stem cells ( hMSCs ) .METHODS: Two recombinant plasmids for over-expression of caspase-8 shRNA, pAd-Cap8 shRNA1 and pAd-Cap8 shRNA2, were constructed.Caspase-8 mRNA was determined in pAd-Cap8 shRNA-transfected human HEK293 cells by Q-PCR.The screened pAd-Cap8 shRNA was used to construct the recombinant adenovirus plasmid , which was linearized and transfected into HEK 293 cells for packaging and amplification of the recombi-nant adenovirus rAd-Cap8 shRNA.The expression of caspase-8 at mRNA and protein levels was determined by Q-PCR and Western blotting .Annexin V/PI staining and determination of caspase-8 activity were performed to assess apoptosis of hM-SCs under the conditions of serum deprivation and hypoxia .The mRNA expression of vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), insulin-like growth factor 1 (IGF-1), Bcl-2 and Bcl-xL was analyzed by Q-PCR.RESULTS:The pAd-Cap8 shRNA, which efficiently inhibited caspase-8 expression, was screened by Q-PCR.The recombinant adenovirus plasmid for caspase-8 shRNA was constructed and used to package and amplify the recombinant ad-enovirus ( rAd)-Cap8 shRNA successfully .rAd-Cap8 shRNA-mediated caspase-8 shRNA markedly inhibited caspase-8 ex-pression in hMSCs .Over-expression of caspase-8 shRNA by infection of rAd-Cap8 shRNA also efficiently decreased the ap-optotic rate and caspase-8 activity in hMSCs under the conditions of serum deprivation and hypoxia , with up-regulation of the mRNA expression of HGF, IGF-1 and Bcl-2.CONCLUSION:Caspase-8 shRNA attenuates hMSC apoptosis under the conditions of serum deprivation and hypoxia .
RESUMEN
Objective To enable medical institutions and personnel in the case of a medical encroachment dispute to provide precise proof in court, use standard format for the legal complaint, employ the appropriate proofgathermg method, ensure the simplicity of the proof-providing procedure, and avoid the waste of manpower and material resources and the liability of economic compensations in the course of compromise settlement, mediation by the health administrative department or adjudication by the court. Methods Regulations were laid down within the hospital for proof-providing procedures, methods, contents, the contents and format of legal complaints and reply letters, and documents in preparation for the provision of proof. Results Through the handling of over 900 medical encroachment disputes, the above methods proved to be practical, applicable and up to the standard. They could save the time of the hospital, the patient and the lawyer on blind surveys, reduce proof-providing time, and improve efficiency in handling medical encroachment disputes. Conclusion Standardizing proof-providing procedures, methods, and contents and the format of legal complaints or reply letters is of great significance in improving efficiency in handling medical encroachment disputes and reducing the hospital's chances of losing lawsuits.