Effects of Quercetin on Acrylamide-Induced Variation of Serum Elements in Rats.

Acrylamide (AA) is an organic chemical widely existing in the public diet, especially in foods with high-temperature fried and baked starchy and may have various adverse health effects on organisms. The purpose of this study was to investigate whether quercetin plays a protective role in AA-induced element variation in rats. Rats were randomly divided into the control group, AA-treated group [5 mg/kg body weight (bw)], two dosages of quercetin-treated groups (10 and 50 mg/kg·bw, respectively), and two dosages of quercetin plus AA-treated groups. After a 16-week treatment, the serum samples of rats were collected. Serum elements were analyzed by using inductively coupled plasma mass spectrometry (ICP-MS) combined with multivariate statistical analysis, and antioxidant indices, lipid peroxidation indicator, as well as inflammatory biomarkers, were also detected. The accuracy and precision of the method were verified, and all the validated data are within the satisfactory range. The results showed that the levels of vanadium (V), copper (Cu), zinc (Zn), selenium (Se), cobalt (Co), and magnesium (Mg) in serum were significantly lower (p < 0.01), while serum calcium (Ca) level was significantly higher (p < 0.01) in AA-treated group compared with the control group. When high-dose quercetin was administered to rats combined with AA, a significant recovered effect for the above elements levels was observed compared with the AA-treated group. This study suggests that quercetin (50 mg/kg·bw) exerts a regulatory and protective role in AA-induced variation of serum elements via reducing oxidative stress and inhibiting inflammation.

Risk factors for acute respiratory distress syndrome in patients with traumatic hemorrhagic shock / 北京大学学报(医学版)

Objective:To investigate the risk factors of acute respiratory distress syndrome(ARDS)after traumatic hemorrhagic shock.Methods:This was a retrospective cohort study of 314 patients with traumatic hemorrhagic shock at Trauma Medicine Center,Peking University People's Hospital from De-cember 2012 to August 2021,including 152 male patients and 162 female patients,with a median age of 63.00(49.75-82.00)years.The demographic data,past medical history,injury assessment,vital signs,laboratory examination and other indicators of these patients during hospitalization were recorded.These patients were divided into two groups,ARDS group(n=89)and non-ARDS group(n=225)ac-cording to whether there was ARDS within 7 d of admission.Risk factors for ARDS were identified using Logistic regression.The C-statistic expressed as a percentage[area under curve(AUC)of the receiver operating characteristic(ROC)curve]was used to assess the discrimination of the model.Results:The incidence of ARDS after traumatic hemorrhagic shock was 28.34％.Finally,Logistic regression model showed that the independent risk factors of ARDS after traumatic hemorrhagic shock included male,histo-ry of coronary heart disease,high acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ)score,road traffic accident and elevated troponin Ⅰ.The OR and 95％confidence intervals(CI)were 4.01(95％CI:1.75-9.20),5.22(95％CI:1.29-21.08),1.07(95％CI:1.02-1.57),2.53(95％CI:1.21-5.28),and 1.26(95％CI:1.02-1.57),respectively;the P values were 0.001,0.020,0.009,0.014,and 0.034,respectively.The ROC curve was used to analyze the value of each risk factor in predicting ARDS.It was found that the AUC for predicting ARDS after traumatic hemor-rhagic shock was 0.59(95％CI:0.51-0.68)formale,0.55(95％CI:0.46-0.64)for history of coronary heart disease,0.65(95％CI:0.57-0.73)for APACHE Ⅱ score,0.58(95％CI:0.50-0.67)for road traffic accident,and 0.73(95％CI:0.66-0.80)for elevated troponin Ⅰ,with an overall predictive value of 0.81(95％CI:0.74-0.88).Conclusion:The incidence of ARDS in pa-tients with traumatic hemorrhagic shock is high,and male,history of coronary heart disease,high APACHE Ⅱ score,road traffic accident and elevated troponin Ⅰ are independent risk factors for ARDS after traumatic hemorrhagic shock.Timely monitoring these indicators is conducive to early detection and treatment of ARDS after traumatic hemorrhagic shock.

Predictive value of pre-surgical 18F-FDG PET/CT metabolic parameters for mediastinal lymph node metastasis in patients with lung adenocarcinoma / 中华核医学与分子影像杂志

Objective:To investigate predictive value of model based on pre-surgical 18F-FDG PET/CT metabolic parameters for mediastinal lymph node metastasis (LNM) in lung adenocarcinoma. Methods:A total of 288 patients with lung adenocarcinoma (135 males, 153 females, age (61.6±8.5) years) who diagnosed and treated in the Fourth Hospital of Hebei Medical University from January 2016 to February 2021 were enrolled retrospectively. All patients underwent 18F-FDG PET/CT examination within 1 month before operation, and underwent complete resection of primary lung tumor and standard lymph node dissection. PET/CT parameters were extracted (PET metabolic parameters: minimum SUV(SUV min), SUV max, SUV mean, SUV standard deviation (SUV std), metabolic tumor volume (MTV) and total lesion glycolysis (TLG); CT parameters: minimum CT value (HU min), maximum CT value (HU max), mean CT value (HU mean), CT value standard deviation (HU std)). Multivariate logistic regression analysis was used for screening parameters and establishing model to predict LNM. ROC curves analyses were used to evaluate the predictive performance of models. Results:Among 288 patients, 90 had LNM, and 361 metastatic lymph nodes (N1: 186, N2: 175) were reported by pathology. SUV min (odds ratio ( OR)=1.859, 95% CI: 1.074-3.220, P=0.027), SUV max ( OR=2.255, 95% CI: 1.306-3.893, P=0.004), SUV mean ( OR=0.277, 95% CI: 0.115-0.665, P=0.004) were predictors of LNM. The AUC of PET/CT model was 0.849 (95% CI: 0.804-0.893), and the sensitivity, specificity, accuracy, and positive and negative predictive values were 87.8%(79/90), 72.2%(143/198), 77.1%(222/288), 59.0%(79/134) and 92.9%(143/154), respectively. Conclusion:The model based on 18F-FDG PET/CT metabolic parameters can improve the accuracy of pre-surgical N-staging in patients with lung adenocarcinoma.

Four-year outcomes of macular buckling for traction maculopathy in highly myopic eyes / 中华眼底病杂志

Objective:To observe the long-term efficacy and safety of macular buckling (MB) in the treatment of high myopia traction maculopathy.Methods:A retrospective clinical study. From January 2014 to December 2017, 57 eyes of 57 patients with high myopia traction maculopathy who underwent MB treatment at Zhongshan Ophthalmic Center of Sun Yat-sen University were included in the study. Among them, there were 15 males with 15 eyes, average age was 51.80±10.72 years; there were 42 females with 42 eyes, average age was 59.14±11.51 years. There were 21 eyes of 21 cases with highly myopic macular hole with macular detachment (MHMD), and 36 eyes in 36 cases with highly myopic foveoschisis with macular detachment (FSMD), and they were grouped accordingly. All patients underwent best corrected visual acuity (BCVA), optical coherence tomography (OCT), and axial length (AL) measurements. The standard logarithmic visual acuity chart was used for BCVA examination, which was converted into logarithm of the minimum angle of resolution (logMAR) visual acuity during statistics. All patients underwent MB, either on its own or combined with vitrectomy. Patients with significant vitreous macular traction on OCT were treated with combined surgery. One, 3, 6 months and 1, 2, 3, and 4 years after the operation, the same equipment and methods before the operation were used to conduct related examinations, and the long-term efficacy and safety of the two groups of eyes were observed.Results:Before surgery, the logMAR BCVA of eyes in MHMD group and FSMD group were 1.35±0.47 and 1.17±0.59, respectively; 4 years after surgery, they were 1.02±0.49 and 0.73±0.55, respectively. The BCVA improved significantly at postoperative 4 years than preoperative in both groups ( P=0.039, 0.001). In the eyes with MHMD, the BCVA was found to be significant improved 3 years after surgery ( P=0.042). Whereas, in the eyes with FSMD, the BCVA was found to be significantly improved 3 months after surgery ( P=0.013). Macular reattachment was achieved in 100% of cases, while macular hole closure rate was achieved in 66.7% in the MHMD group. In the FSMD group, either macular reattachment rate or the foveoschisis resolution rate was 97.2%. After surgery, choroidal neovascularization was observed in 2 eyes, and 3 eyes with intraretinal cyst. Conclusion:MB may represent a safe and effective surgical option for the treatment of high myopia maculopathy.

Discriminant analysis of the risk of nosocomial mortality in patients with traumatic hemorrhagic shock / 中华普通外科杂志

Objective:To investigate the use of discriminant analysis to predict the risk of nosocomial mortality in patients with traumatic hemorrhagic shock.Methods:The clinical data of 238 patients with traumatic hemorrhagic shock admitted to Peking University People's Hospital from Sep 2013 to Aug 2020 were retrospectively analyzed. Patients were divided into survival group (214 cases) and death group (24 cases). Stepwise discriminant analysis was used to establish a discriminant model.Results:The difference of history of stroke (9.8% vs. 25.0%), main site of bleeding (extremities)(58.9% vs. 29.2%), APACHEⅡ score (16.4±5.1 vs. 23.2±6.1), blood lactic acid [2.1(1.1-3.5) mmol/L vs. 4.9(2.0-13.4) mmol/L] and surgery (92.5% vs. 58.3%) between the two groups was all statistically significant (all P<0.05). Finally, There are five indicators that entered the discriminant model: history of stroke, main site of bleeding (extremities), blood lactic acid, APACHE Ⅱ score and surgery. The area under the ROC curve for predicting the risk of mortality in patients with traumatic hemorrhagic shock was 0.857, 95% CI 0.754-0.959. Conclusions:The established discriminant model has a high accuracy in predicting the risk of in-hospital mortality in patients with traumatic hemorrhagic shock.

Study of occult uterine perforation in brachytherapy of cervical cancer / 中华放射肿瘤学杂志

Objective:To evaluate the difference of dosimetry between three-dimensional and two-dimensional plans based on CT images of occult perforation in brachytherapy of cervical cancer, aiming to provide clinical reference.Methods:A total of 817 patients with cervical cancer received simple intrauterine (intrauterine tandem plus vaginal colpostats) three-dimensional brachytherapy in Chongqing University Cancer Hospital from January 2019 to December 2020 were retrospectively reviewed. Among them, 16 patients had occul uterine perforation. Based on Oncentra Brachy Therapy plan system, the single prescription dose was 6Gy. Three-dimensional (3D group) and two-dimensional (2D group) plans were designed on the perforated CT images The target volume, conformal index (CI), conformal index coformity index (COIN) and organs-at-risk (OAR) D 2cm 3 parameters were used to assess the plans between two groups. Results:The incidence of pccult uterine perforation was 1.96%(16/817) during brachytherapy for cervical cancer. The volume of prescription dose curve in the 3D group was (40.74±14.98) cm 3, significantly smaller compared with (91.46±19.71) cm 3 in the 2D group ( P<0.05), whereas the volume of the high-risk clinical target area wrapped by prescription dose curve did not significantly differ between two groups ( P>0.05). The CI and COIN in the 3D group were 0.79±0.10 and 0.72±0.96, significantly higher compared with 0.38±0.09 and 0.37±0.18 in the 2D group (both P<0.05). The D 2cm 3 of bladder, rectum, sigmoid colon, small intestine in the 3D group were (306.06±77.57) cGy, (252.27±72.60) cGy, (127.25±62.84) cGy and (228.79±94.90) cGy, significantly lower than (548.03±164.21) cGy, (411.16±118.74) cGy, (227.45±94.48) cGy and (450.95±157.96) cGy in the 2D group (all P<0.05). Conclusions:Application of image guidance in brachytherapy of cervical cancer is helpful to detect occult uterine perforation. When occult uterine perforation occurs, the use of three-dimensional plan can basically meet the clinical needs, which is significantly better than the two-dimensional plan.

Risk factors associated with myocardial injury after non-cardiac major surgery in ICU admitted patients / 中华普通外科杂志

Objective To analyze the risk factors related to the myocardial injury after non-cardiac surgery (MINS) in patients who underwent major abdominal surgery.Methods The clinical data of all patients admitted in the surgical ICU of Peking University People's Hospital from Jan 2016 to Dec 2018 were analyzed.Logistic multivariate analysis was performed to analyze the association of clinical characteristics with the incidence of MINS.Results A total of 322 patients were included,48.4％ (156/322) were diagnosed as with MINS.97.4％ (152/156) of MINS occurred during the first 72 h of admission.Multivariate analysis showed that independent predictive factors of MINS were age ＞ 65y (OR =1.747,P =0.021),body mass index (OR =1.085,P =0.008),acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ,OR =1.066;P =0.047),hypertension (OR =1.747,P =0.027),blood lactate (OR =1.393,P=0.001) and acute kidney injury (OR=2.065,P=0.047).Conclusions The incidence ofMINS in patients who underwent major abdominal surgery was high.Age,body mass index,APACHE Ⅱ score,hypertension,blood lactate level and acute kidney injury after surgery were independent risk factors of MINS.

Risk factors associated with myocardial injury after non-cardiac major surgery in ICU admitted patients / 中华普通外科杂志

Objective@#To analyze the risk factors related to the myocardial injury after non-cardiac surgery (MINS) in patients who underwent major abdominal surgery.@*Methods@#The clinical data of all patients admitted in the surgical ICU of Peking University People′s Hospital from Jan 2016 to Dec 2018 were analyzed. Logistic multivariate analysis was performed to analyze the association of clinical characteristics with the incidence of MINS.@*Results@#A total of 322 patients were included, 48.4% (156/322) were diagnosed as with MINS. 97.4% (152/156) of MINS occurred during the first 72 h of admission. Multivariate analysis showed that independent predictive factors of MINS were age >65y (OR=1.747, P=0.021), body mass index (OR=1.085, P=0.008), acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ, OR=1.066; P=0.047), hypertension (OR=1.747, P=0.027), blood lactate (OR=1.393, P=0.001) and acute kidney injury (OR=2.065, P=0.047).@*Conclusions@#The incidence of MINS in patients who underwent major abdominal surgery was high. Age, body mass index, APACHE Ⅱ score, hypertension, blood lactate level and acute kidney injury after surgery were independent risk factors of MINS.

CRISPR-Cas9-based site-directed knock-in of VEGF165 gene in a HEK293T cell / 国际生物医学工程杂志

Objective To construct a human renal epithelial cell line HEK293T by CRISPR-Cas9-based site-directed knock-in of vascular endothelial growth factor 165 (VEGF165) gene, and avoid the off-target effect caused by lentivirus infection. Methods The VEGF165 expression vector with homologous arm (pUCm-T-VEGF165 plasmid) and the sgRNA expression vector [pSpCas9(BB)-2A-Puro-sgRNA plasmid] were designed and constructed based on the DNA sequence of the EZH2 gene, and then co-transfected into HEK293T cells. The expression of VEGF165 mRNA was detected by qPCR and the expressions of VEGF165 proteins were detected by Western Blot. Results The qPCR and Western Blot results showed that, comparing with the control, the pUCm-T-VEGF165 plasmid and pSpCas9(BB)-2A-Puro-sgRNA plasmid, the expression of the co-transfection plasmid were significantly increased, i.e. 3.42±0.30 vs. 1.02±0.21, 1.13±0.16 and 0.98±0.18 for the VEGF165 mRNA level (all P<0.01), and 1.13±0.16 vs. 1.02±0.06, 0.88±0.03 and 0.80±0.05 for the VEGF165 protein level (all P<0.01), respectively. Besides, the expression of EZH2 was significantly down-regulated, i.e. 0.14±0.06 vs. 1.08±0.11, 1.02±0.12 and 1.13±0.16 for the EZH2 mRNA level (all P<0.01), and 0.23±0.03 vs. 1.05±0.13, 0.91±0.04 and 0.81±0.06 for the EZH2 protein level (all P<0.01), respectively. This result showed that the VEGF165 was successfully inserted into the EZH2 genome, interfering the EZH2 expression. Conclusions VEGF165 gene can be successfully knocked into HEK293T cells by CRISPR/Cas9 system.

New progress of nutrition for severe patients at home and abroad / 中国中西医结合急救杂志

In recent years, the researches on nutrition for critically ill patients had advanced quickly, especially the domestic severe illness nutritional studies. In this review, the risk of malnutrition in severe patients and the pros and cons of nutritional evaluation stool, illustration of the use of low calorie nutrition in critically ill patients, the comparison of the advantage and disadvantage between enteral nutrition and parenteral nutrition, and the immune nutritional method for treatment of critically ill patients were explained; the key point of this summary is on the advance of severe illness nutrition at home compared with that at abroad in order to make a suggestion to direct the future development of clinical nutrition.

Construction and identification of LSD1 overexpression and demethylation disfunction plasmids / 国际生物医学工程杂志

Objective To construct rat lysine-specific histone demethylase 1 (LSD1) overexpression plasmids and LSD 1 demethylation fragment disfunction plasmids,and to evaluate their expression levels in HEK293T cells.Methods LSD1 fragments were amplified by PCR,and LSD1 demethylation disfunction fragments were amplified by overlap PCR.Rat-specific LSD1 overexpression plasmids and LSD1 demethylation disfunction plasmids were constructed and verified by agarose gel electrophoresis and sequencing.HEK293T cells were infected using the validated recombinant plasmids and blank vectors,and the stably expressed cells were selected by puromycin.The expression of LSD1 in the stably expressed HEK293T was detected by Western Blot and real-time quantitative PCR.Results The results of agarose gel electrophoresis and sequencing showed that LSD1 overexpression plasmids and LSD1 demethylation disfunction plasmids were successfully constructed.The Western Blot and real-time quantitative PCR results showed that compared with the blank group,the relative expression of LSD1 and mRNA in the LSD1 overexpression group and the LSD1 demethylation disfunction group were up-regulated,and the differences were statistically significant(all P＜0.01).Conclusions The constructed LSD1 overexpression plasmids and LSD1 demethylationi disfunction plasmids can achieve overexpression of LSD1 gene in HEK293T cells.This paper lays a foundation for further study of the relationship between LSD 1 gene and related diseases and demethylation of LSD 1.

Analysis of the anxiety situation and the influencing factors among gerontal patients with osteoporosis / 中国基层医药

Objective To understand the anxiety among gerontal patients with osteoporosis,and to analyze the correlated factors of the anxiety.Methods The social information of the gerontal patients with osteoporosis were investigated,as well as by SAS for the anxiety,and by NRS,PSQI,AHSMSRS for their pain level,sleep quality and healthy management.Results The incidence rate of anxiety was 20.3%,and the average score was (41 .1 4 ± 9.71 )points.Some factors had influence upon the rate,which included sex,month income,sleep quality,self -care ability,pain level,self -management and exercise.Multivariate logistic regression analysis showed that exercise and pain level were the major risk factors.The differences were statistically significant(t =3.033,2.727,P =0.003, 0.007).Conclusion It's higher that the incidence rate of anxiety among gerontal patients with osteoporosis maybe influenced by species of factors,and the quality of life among them needs to be improved by intervention clinically implicated in those factors.

Construction of EZH2 gene site-directed knock-in Hut78 cells by CRISPR/Cas9 system / 天津医药

Objective To construct the Hut78 cell line with EZH2 gene knocked into by CRISPR/Cas9 system. Methods The EZH2 expression vector pMD-18T-EZH2 with homologous arm and the sgRNA expression vector pSpCas9 (BB)-2A-Puro-sgRNA, which could cut the double stranded genomic DNA, were constructed, and the two vectors were co-transfected into Hut78 cells. Then the expression of EZH2 mRNA was detected by qPCR, and the expressions of EZH2 and H3K27me3 proteins were detected by Western blot assay. Results The pMD-18T-EZH2 and pSpCas9(BB)-2A-Puro-sgRNA recombinant vectors were confirmed by DNA sequencing. When Hut78 cells were transfected with the two recombinant plasmid, qPCR results showed that the expression of EZH2 mRNA was significantly increased, and Western blot analysis showed that the expressions of EZH2 and H3K27me3 proteins were significantly increased. Conclusion EZH2 gene is successfully knocked into Hut78 cells by CRISPR/Cas9 system.

The role and mechanism of resveratrol on trabecular meshwork cells induced by H2O2 and TGF-β2 / 天津医药

Objective To investigate hydrogen peroxide (H2O2) and transforming growth factor-β2 (TGF-β2) induced fibronectin (FN), collagen 1 (COL1), nuclear factor (NF)-κB P65 proteins and interlukin (IL)-1βgene expression in human trabecular meshwork cells (HTMCs), and the interventional mechanism of resveratrol (RSV). Methods (1) HTMCs with 70 to 80%confluency were divided into 5 groups. The experimental groups were treated with serum-free medium and with H2O2 at concentrations of 150, 300, 450 and 800μmol/L. The control group was treated with 0μmol/L H2O2. The protein levels of FN, COL1, NF-κB P65 and NF-κB P65 phosphorylation (P-NF-κB P65) were measured by Western blot assay. The expression of IL-1βgene was measured by qPCR. (2) HTMCs were divided into 3 groups. The control group was treated withserum-free medium and without H2O2 and RSV. The H2O2 group was treated with 300μmol/L H2O2. The H2O2+RSV group was treated with 300μmol/L H2O2 and 25μmol/L resveratrol (RSV). The expressions of proteins and genes mentioned above were detected in three groups. NF-κB P65 nuclear translocation was assessed by immunofluorescence technique. (3) HTMCs were divided into 3 groups. The control group was treated with serum-free medium and without TGF-β2 and RSV. The TGF-β2 group was treated with 5μg/L TGF-β2. The TGF-β2+RSV group was treated with 5μg/L TGF-β2 and 25μmol/L RSV. The expressions of proteins and genes mentioned above were detected in three groups. Results (1) Compared with control group, the protein levels of FN and P-NF-κB P65 were significantly increased in 150, 300, 450 and 800μmol/L groups,the expression levels of COL1 protein and IL-1β gene were significantly increased in 300, 450 and 800 μmol/L groups (P <0.05). There were no statistical significances between other indicators. (2) The expression levels of FN, COL1, P-NF-κB P65 proteins and IL-1βgene were significantly higher in H2O2 group than those in control group, and which were significantly lower in H2O2+RSV group than those in H2O2 group. Compared with control group, only the expression of IL-1βgene was decreased in H2O2+RSV group (P < 0.05). NF-κB P65 was only expressed in cytoplasm in control group, while it was expressed in both cytoplasm and nucleus in H2O2 group. Compared with H2O2 group, NF-κB P65 was mainly expressed in cytoplasm. (3) Compared with control group, the expressions of FN, COL1, P-NF-κB P65 proteins and IL-1β gene were significantly increased in TGF-β2 group (P < 0.05). Compared with TGF-β2 group, the indicators mentioned above were significantly decreased in TGF-β2+RSV group (P<0.05). Conclusion H2O2 and TGF-β2 can upregulate the expression of FN, COL1, P-NF-κB P65 proteins and IL-1βgene in HTMCs, which may be involved in the development and progression of glaucoma. RSV can inhibit the influence of H2O2 and TGF-β2 in HTMCs and exert a protective effect on glaucoma.

Study on p21 gene knock out in G401 cell line by using CRISPR/Cas9 system / 天津医药

Objective To knock out p21 gene in human malignant rhab doid tumor(MRT)cell line G401 by using CRISPR/Cas9 genome engineering technology. Methods The expression of p21 was detected by reverse transcription quantitative PCR (RT-qPCR) and Western blot assay in several MRT cell lines. The guide RNA was designed by targeting the third exon of p21 gene,which encoded its home domains, and then subcloned into lentiCRISPR v2 vector and validated sequencing. The validated plasmids were further used to package and produce the lentivirus in 293T cells, and the G401 cells were infected, then puromycin was used to screen positive cells, and the clusters of G401 monoclonal cells, were obtained by selecting monoclonal cells and culturing under the microscope. The RNA and protein of new clonal cell line were extracted, and RT-qPCR and Western blot assay were applied to confirm whether p21 was successfully knocked out. Results The p21 was highly expressed in MRT tumor cells. The CRISPR/Cas9 lentivirus plasmids, targeted p21 gene were successfully constructed. Compared with negative control group,the expression of p21 was not detected in G401 monoclonal cells, which were successfully screened. Conclusion In view of the difficult transfection of cells such as G401, p21 knockout stable cell line has been successfully constructed by using CRISPR/Cas9 system, which lays the foundation for further study of the mechanism of p21 in MRT tumors .

Construction of Asxl2 gene knock out stable NIH3T3 cell line with CRISPR/Cas9n system / 天津医药

Objective To knock out Asxl2 gene in murine embryonic fibroblast cell line NIH3T3 using CRISPR/Cas9n system. Methods A pair of sgRNAs which targeted exon 5 of Asxl2 gene were designed and subcloned into the pX462 vec?tor. The recombined plasmids were verified by sequencing and transfected into NIH3T3 cell line. Single cells were isolated through serial dilutions, followed by an expansion period to obtain new monoclonal cell lines. The genomic DNA of the new monoclonal cell lines was extracted and a DNA fragment flanked the target site was amplified by genotyping PCR then se?quenced. Lastly, western blotting were applied to confirm whether Asxl2 was successfully knocked out. Results The CRIS?PR/Cas9n plasmids that targeted Asxl2 were successfully constructed. NIH3T3 cells were co-transfected with the two recom?binant constructs. After puromycin selection, subclonal cell lines were obtained and one of them was validated by genotyping PCR-sequencing. Western blotting also confirmed that Asxl2 was completely depleted in the NIH3T3 cell line. Conclu?sion CRISPR/Cas9n plasmids that targeted Asxl2 were successfully constructed therefore a Asxl2 knockout NIH3T3 stable cell line was established via this system.

Risk factors of major adverse cardiac events of critical care patients after non cardiac surgery / 中华普通外科杂志

Objective To investigate the incidence and the risk factors of major adverse cardiac events of critical care patients after noncardiac surgery.To study the incidence of myocardial injury after noncardiac surgery (MINS).Methods A retrospective analysis of critical care patients (n =1 087) after noncardiac surgery from January 2012 to January 2013 in our hospital was carried out.The clinical data of the medical history, intraoperative conditions, postoperative conditions and cardiac troponin Ⅰ (cTNI) were collected.Major adverse cardiac events included unstable angina, non fatal myocardial infarction, severe arrhythmia, heart failure and cardiovascular death.Risk factors of major adverse cardiac events of critical care patients after noncardiac surgery were analyzed using Logistic regression.Results The 30 d non cardiac surgery incidence of major adverse cardiac events was (94/1 087) 8.6％, the incidence of MINS was (168/1 087) 15.5％, Logistic regression analysis showed that the risk factors of major adverse cardiac events after non cardiac surgery were age (OR 1.03,95％ CI 1.01-1.05, P =0.002), a history of chronic renal insufficiency(OR 3.12,95％ CI 1.44-6.74,P =0.004), rise of cTNI 24 h after operation (OR 2.04,95％ CI 1.16-3.58,P =0.014) and use of vasopressor drugs within 24 h after operation (OR 2.34,95％ CI 1.25-4.38,P =0.008).Conclusions The incidence of major adverse cardiac events and the MINS of critical care patients after noncardiac surgery is high.Old age, history of chronic renal insufficiency, rise of cTNI 24 h after operation and vasopressor drugs within 24 h after operation are the independent risk factors.

Investigation on the caring behaviors of family caregivers of patients of indwelling catheter / 重庆医学

Objective To investigate the family caregivers of patients with indwelling catheter care behavior .By analyzed its in‐fluencing factors to help caregivers improve the quality of life for patients .Methods Convenience sampling method by visit 70 pri‐mary caregivers of family indwelling catheter for the patient′s .They were investigated with gathering general information and Gen‐eral Self‐Efficacy Scale(GSES) ,following with caregivers awareness questionnaire indwelling catheter .Indwelling catheter care be‐havior questionnaire survey analyze behavior and other items to take care of the relationship .Results The general self‐efficacy and caregiver indwelling catheter cognition ,which account for 73 .9% of the total variance of caring behaviors ,were the important fac‐tors affecting care behaviors .Conclusion There is a big shortage in indwelling catheter home care caregiver knowledge and care be‐haviors .According to above professionals can provide targeted guidance for caregivers .

Retroperitoneal Hodgkin's lymphoma involving right ureter misdiagnosed as right ureteral carcinoma with lymphadenopathy: a case report and review of literature / 中华泌尿外科杂志

Objective To investigate the clinical and pathological features,diagnosis,differential diagnosis,treatment and prognosis of secondary Hodgkin's lymphoma of ureter.Methods We retrospectively analyzed one case with retroperitoneal Hodgkin's lymphoma involving right ureter.A 63-year-old male patient was referred to our hospital on January 22th,2013 with right hydronephrosis for 15 days,detected by abdominal color Doppler ultrasound due to constipation and difficult defecation without any urinary symptoms.Urinary CT scan revealed that there were dilation and hydrops of right pelvic and ureter in the middle and upper segment.The distal portion of ureteral wall was thick with a stenosis lumen.There was multiple larged lymph nodes surrounding the abdominal aorta.So,it was clinically diagnosed as right ureteral carcinoma with lymphadenopathy and right hydronephrosis.The patient was then performed ureteroscopic inspection under general anesthesia.There was too narrow to go through smoothly the proximal ureter when the ureteroscope ascended 15 cm.Then,the exploratory surgery of right ureter was subsequently undergone.During operation,periureteral nodular mass and thickened wall were seen in the distal segment of ureter.Then the ureter lesion was partially excised with 1 cm and fast frozen pathology was undergone during operation.The result suggested chronic inflammation of ureteral.A 6Fr double J stent was left in the ureter.Then,the ureter was closed by 3-0 monocryl suture.Results The final pathological diagnosis was retroperitoneal Hodgkin's lymphoma with the lymph node involving the right ureter.After 40 days,the patient manifested high fever,night sweat,general lymphoadenomegalies.So,the clinical stage was classified as stage Ⅱ EB and poor prognosis group.Then,the patient received regular chemotherapy according to the ABVD (pirarubicin 20 mg/m2,bleomycin 10 mg/m2,vinblastine 1.4 mg/m2,dacarbazine 375 mg/m2) regimen over 8 cycles.Finally the symptomatology and diagnostic reassessment (hemato-chemical examinations and PET/CT) showed a clinical complete recovery after the follow-up period of 3 months.The follow-up plan is ongoing.Conclusions Because secondary Hodgkin's lymphoma of ureter is rare with concealed onset feature,lacking of characteristic clinical manifestations,it is relatively difficult to achieve the diagnosis and differential diagnosis.Therefore,pathology and immunohistochemistry should be assigned to confirm the diagnosis.It is stressed that physicians should be aware of the possibility of malignant lymphoma when unexplained hydronephrosis and thickness of the ureteral wall are found clinically.Generally it has a good prognosis and combination therapy for the treatment is given priority to with chemotherapy.

Early compliance and efficacy of sublingual immunotherapy in patients with allergic rhinitis / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#The study aimed to investigate the efficacy and adverse effects of sublingual immunotherapy (SLIT) of dust mite drops to allergic rhinitis with mite allergy. The compliance and satisfaction of SLIT were also assessed.@*METHOD@#One hundred and three patients of allergic rhinitis sensitive to dust mites were treated with SLIT for 6 months or more. The symptom questionnaire,including items on rhinorrhea, sneezing, nasal obstruction, itchy nose, olfactory disturbance, eye discomfort and sleep disturbance were obtained before and 6 months after SLIT. The patients' satisfaction and adverse effects were also investigated.@*RESULT@#Seventy-five of the 103 patients insist on SLIT for more than 6 months and completed the questionnaire. The duration of receiving SLIT was 9.8 months on average (range from 6 to 13 months). The satisfaction rate was 89.3%. The drop-out rate of SLIT was 31.0%.@*CONCLUSION@#The subjective symptoms were improved with SLIT in patients with allergic rhinitis sensitive to dust mites. The drop out rate was high despite of the symptomatic improvement.