Preparation of colloidal gold test strips for the detection of antibodies to peste des petits ruminants based on monoclonal antibodies to N protein / 生物工程学报

A simple, fast, and visual method for detecting antibodies against peste des petits ruminants virus (PPRV) using colloidal gold strips was developed. In this study, the pET-32a-N was transformed into Escherichia coli Rosetta (DE3) for expression. Hybridoma cell lines were generated by fusing SP2/0 myeloma cells with splenocytes from immunized mice with the expressed and purified N protein of PPRV. The PPRV N protein was labeled with colloidal gold particles as the gold-labeled antigen. The N protein served as the gold standard antigen and as the test (T) line-coated antigen, while the monoclonal antibody served as the quality control (C) line-coated antibody to assemble the colloidal gold immunochromatographic test strips for detecting antibodies against the N protein of PPRV. Hybridoma cell line designated as 1F1 was able to stably secrete the monoclonal antibody against the N protein of PPRV. The titer of 1F1 monoclonal antibody in ascites was 1:128 000 determined by indirect enzyme-linked immunosorbent assays (ELISA), and the immunoglobulin subtype of the monoclonal antibody was IgG1, with kappa chain. The obtained monoclonal antibody was able to specifically recognize the N protein of PPRV, as shown by Western blotting and indirect immunofluorescent assay (IFA). The developed colloidal gold test strip method was able to detect PPRV antibodies specifically, and there was no difference between different batches of the test strips. Testing of a total of 122 clinical sera showed that the compliance rate of the test strip with ELISA test was 97.6%.The test strip assay developed in this study has good specificity, reproducibility, and sensitivity, and it can be used for the rapid detection of PPRV antibodies.

Case report of congenital disorder of glycosylation type Ia combined with dilated cardiomyopathy / 中华实用儿科临床杂志

Clinical data and follow-up of a case of congenital disorder of glycosylation type Ia (CDG-Ia) combined with dilated cardiomyopathy admitted to the Department of Cardiology, Children′s Hospital of Nanjing Medical University were analyzed retrospectively.The 5-year-old female patient was admitted in December 2016 due to recu-rrent shortness of breath for 2 months.Clinical symptoms and signs included repeated attacks of shortness of breath, physical retardation, malnutrition, binocular esotropia, multiple episodes of hypoglycemia, hepatosplenomegaly, hypotonia and other multi-system damages.Cardiac echocardiography suggested the feature of dilated cardiomyopathy, including the significant enlargement of the left ventricle, and decreased systolic function.Genetic testing revealed a compound heterozygous mutation in the PMM2 gene, and as a result, the patient was diagnosed as CDG-Ia.The patient′s condition improved after symptomatic treatments such as Cedilanid, Dopamine, Dobutamine, Furosemide, as well as support treatments like myocardium nutrition, blood sugar maintenance, liver protection, etc.After discharge, the patient was given oral Digoxin, Betaloc, Captopril and diuretics, and hypoglycemia-controlling agents.The patient was followed up every 3-6 months.After more than 2 years of follow-up, the heart function and heart enlargement gradually returned to normal.During the Corona Virus Disease 2019 outbreak, self-withdrawal continued for 2 months.Re-examinations showed decreased cardiac function and enlarged left ventricle again.Medications were resumed again, and the patient was followed up closely.This case report suggested that CDG-Ia may be associated with dilated cardiomyopathy, and the cardiac phenotype may be improved by symptomatic supportive treatment.

Risk factors and prediction model of thrombocytopenia in children with patent ductus arteriosus after transca-theter closure / 中华实用儿科临床杂志

Objective:To explore the risk factors of thrombocytopenia in children with patent ductus arteriosus (PDA) after transcatheter closure, and to establish a prediction model of thrombocytopenia after transcatheter closure of PDA.Methods:A total of 39 PDA children with thrombocytopenia after transcatheter closure treated in Children′s Hospital of Nanjing Medical University from November 2016 to January 2020 were selected.During the same period, 138 PDA children without thrombocytopenia after transcatheter closure were included in the control group. Logistic regression model was used to explore the possible risk factors of thrombocytopenia after transcatheter closure in PDA children, and a random forest model was established to predict the occurrence of thrombocytopenia after transcatheter closure of PDA. Results:After transcatheter closure of PDA, children developed thrombocytopenia within 1 to 7 days, and the platelet count recovered within 2 to 22 days. Logistic regression model suggested that the diameter of pulmonary artery end of arterial catheter ( OR=9.54, 95% CI: 2.08-48.84, P=0.004)and preoperative platelet count( OR=0.99, 95% CI: 0.98-0.99, P=0.001)were correlated with the occurrence of thrombocytopenia after transcatheter closure of PDA.The random forest model indicated that PDA inner diameter was the most important factor for predicting the occurrence of thrombocytopenia after transcatheter closure of PDA. Conclusions:A large diameter of arterial duct is an important risk factor and increased preoperative platelet count is a protective factor for thrombocytopenia after transcatheter closure of PDA.Diameter is of the greatest significance in predicting the occurrence of thrombocytopenia after transcatheter closure of PDA.

Application of rhodamine-based fluorescent molecular probes in visualization of cellular pyruvic acid / 中国药科大学学报

In order to conduct the visualized research of cellular pyruvic acid(PA),several rhodamine-based probes were designed to react with PA by a Schiff-base reaction in aqueous media and exhibit strong fluorescence emission and a pink color owing to the ring-opening of the corresponding spirolactam.The results showed that R6G-NH2had a highest sensitive response to PA with a linearly relationship between the enhancement of fluores-cence intensity at 565 nm and the concentration of PA from 0.2 μmol/L to 120 μmol/L.According to the princi-ple of 3σ,a limit of detection was obtained as low as 0.1 μmol/L.Furthermore,the response of R6G-NH2to PA was hardly affected by conventional biomolecules and ionic species.Therefore,R6G-NH2was successfully used in fluorescence imaging of exogenous PA in non-small cell lung cancer cell A549 and renal epithelial cell 293T,and endogenous PA induced by bisphenol A in epithelial cell 293T.

Exploration of enzyme-MnO2 nanosheets hybridization probe for sensitively colorimetric self-indicating of glucose / 中国药科大学学报

A colorimetric self-indicating probe for glucose was constructed by self-assembly of MnO2 nanosheets (MnO2 NSs) and glucose oxidase(GOD) in this paper.Under the weak acidic medium,glucose oxidase specifically catalyzes glucose into gluconic acid and hydrogen peroxide.The by-product of hydrogen peroxide could efficiently dissolve the MnO2 nanosheets,resulting into a significant decrease of the characteristic absorbance at 374 nm assigned to MnO2 NSs.Furthermore,the absorbance difference was linearly proportional to the concentration of glucose ranging from 1 to 20 μmol/L The fitted curve could be used for quantification of glucose with a correlation coefficient of 0.990 1.And the detection limit as low as 0.1 μmol/L could be reached based on the definition of three times of the deviation of the blank signal (3σ) and there was negligible interference with other co-existing amino acids,anions,cations and protein,which indicated high sensitivity and selectivity of the hybrid probe.The construction strategy of designated probe is readily generalized in principle for detection of numerous analytes in view of reactive property of MnO2 and the diversity of enzymes.

Effect of ultra-early stent-assisted coil embolization on ruptured intracranial aneurysms / 中国脑血管病杂志

Objective To investigate the therapeutic effect of the ultra-early stent-assisted coil embolization of the ruptured intracranial aneurysms. Methods The clinical data of 13 patients with ruptured intracranial aneurysm treated by ultra-early stent-assisted coil embolization were analyzed retrospectively. The preoperative Hunt-Hess gradeⅠ-Ⅱ was in 7 cases,gradeⅢ was in 4 cases,and grade Ⅳ was in 2 cases. The patients were treated with stent-assisted coil embolization under the general anesthesia with endotracheal intubation within 24 h of aneurysm rupture. The postoperative embolization was assessed according to the Raymond grading standard. The postoperative complications and the assessment of the follow-up results from 1 to 6 months after procedure according to the modified Rankin scale (mRS ) scores were observed. Results All 11 patients recovered well,1 case had postoperative hemiplegia,1 case had postoperative bleeding,and none of them died. During the follow-up period,no patients had rebleeding, 1 had recurrence,and DSA revealed that the patient was embolized completely at 2 months after reembolization. Conclusion The method of ultra-early stent-assisted coil embolization of ruptured intracranial aneurysms is feasible. It may improve the cure rate of the ruptured aneurysms and improve the prognosis of patients.

Study on Intelligent Automatic Tracking Radiation Protection Curtain / 中国医疗器械杂志

In order to overcome the shortcomings of traditional X-ray inspection taking passive protection mode, this paper combines the automatic control technology, puts forward a kind of active protection X-ray equipment. The device of automatic detection of patients receiving X-ray irradiation part, intelligent adjustment in patients and shooting device between automatic tracking radiation protection device height. The device has the advantages of automatic adjustment, anti-radiation device, reduce the height of non-irradiated area X-ray radiation and improve the work efficiency. Testing by the professional organization, the device can decrease more than 90% of X-ray dose for patients with non-irradiated area.

Adaptive thermogenesis of the brown adipose tissue in tree shrews (Tupaia belangeri) during cold acclimation / 中国实验动物学报

Objective To investigate the effect of ambient temperature on body mass, thermogenic activity and un-coupling protein-1 ( UCP1) content of brown adipose tissue ( BAT) in tree shrews ( Tupaia belangeri) , and to provide the-oretical basis for establishing tree shrews model of obesity.Methods Forty healthy adult tree shrews with similar body mass were uesd in our experiment.The tree shrews were divided into five groups (n=8):control group (0 d), the ani-mals were maintained under 25 ±1℃ and 12L:12D ( light : dark, lights on 08:00) photoperiod; and the animals were maintained under 5 ±1℃and 12L:12D photoperiod for 7 d, 14 d, 21 d and 28 d groups, respectively.At the end of ex-periment, the changes of body mass, nonshivering thermogenesis (NST), BAT mass and uncoupling protein 1 (UCP1) con-tent were determined.Results Compared with the control group (0 d), the body mass, NST, BAT mass and UCP1 con-tent of the cold acclimation groups were improved significantly, the BAT color also obviously deepened, and after cold accli-mation for 28 d, the body mass, NST, BAT mass and UCP1 content were increased by 26.32%, 20.65, 53.85%and 43%, respectively.Apparently, the UCP1 content was significantly positively correlated with BAT mass and NST.Conclusions BAT proliferation may be induced and UCP1 expression upregulated by cold acclimation in Tupaia belangeri, therefore, en-hancing the thermogenic activity of brown adipose tissue to increase energy expenditure.We would speculate that BAT might be used as a target organ for treatment of obesity by energetic approach in the future.

Synthesis and application of a fluorescent molecular probe for rapid detection of sulfur dioxide residues in traditional Chinese herbs / 中国药科大学学报

@#A fluorescence method based on a “turn-on” rhodamine derivative fluorescent probe was developed for the detection of hydrogen sulfite ions and sulfur dioxide residues in sulfur-fumigated herbs. This method was realized through a well-known aldehyde-bisulfite addition reaction accompanied by a ring opening of spirolactam of probe molecule in pH 4. 8 aqueous ethanol media which resulted in a significant fluorescence and color change. The fluorescence enhancement was linearly proportional to the concentrations of bisulfite ranging from 0. 005 to 20 μmol/L with a correlation coefficient of 0. 998 5. The detection limit was low as to 2. 0 nmol/L and there was no interference with other familiar co-existing anions, cations and reducing reagents indicating high sensitivity and selectivity of the proposed method. In addition, the novel probe was successfully applied for the identification of sulfur dioxide residues in herbs with a satisfactory recovery in five real samples.

Arthroscopy-assisted treatment of severe comminuted distal radial fracture using external fixators / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the outcomes of arthroscopy-assisted treatment of severe comminuted distal radial fracture with external fixators and kirschner wire fixation.</p><p><b>METHODS</b>Twenty-seven cases of severe comminuted distal radial fracture treated between March, 2010 and January, 2012 were reviewed. During the operation, the carpal joint space was expanded with the external fixator, and the fracture was fixed by Kirschner wire after open reduction. The carpal joint was observed intraoperatively with arthroscopy to ensure full reduction, and the distal posterior interosseous nerve was then severed. The results of postoperative X-ray and wrist functional status of the carpal joints were recorded. Another 27 cases of severe comminuted distal radial fracture treated by conventional surgical approach served as the control group.</p><p><b>RESULTS</b>The patients were followed up for a mean of 13.2 (5-27) months. Compared with the conventional surgical approach, arthroscopy-assisted treatment resulted in a significantly shorter operative time with better appearance of the articular surface and also better wrist function assessed using the Krimmer system (P<0.05).</p><p><b>CONCLUSION</b>Arthroscopy-assisted external fixator treatment is effective for management of severe comminuted distal radial fracture and avoids the stair-like appearance of the articular surface to achieve the maximal functional recovery of the carpal joints and reduce traumatic arthritis.</p>

Clinical epidemioiogical analysis of hospital-associated pneumonia in senile patiens in Guangzhou strict / 中国基层医药

Objective To determine the distribution and antibiotic resistance of hospital - associated pneu-monia(HAP) in senile patient,and the high risk factors of HAP,pathogenic bacterium and prognosis,so as to instruct the clinic prevention and treatment. Methods Patients with age over 60 years old,who were diagnosed of HAP with confirmed pathogens. Pathogens were identified, then the antibiotic resistance was determined by Kirty-Baucer disk dif-fusion assay. High risk factors of HAP, pathogenic bacterium and prognosis were analyzed by Logistic regression analy-sis. Results The first 10th pathogens of HAP patients were pseudomonas aeruginosa( 16.1% ),staphylococcus au-reus( 14.6% ) ,klebsiella pneumonia( 10.2% ), escherichia coil (8.8%) ,staphylococcus hemolyticus (7.3%). MR-SA accounted for 90% in staphylococcus aureus. The drug resistance of pseudomonas aeruginosa to imipenem was 29.5%. The high risk factors of HAP were acute brain accident, hypoalbuminemia, tracheal intubation or mechanical ventilation, rennal failure,COPD, gastric tube, anaemia,liver disfunction,long length of stay ( all P < 0.05 ). The mor-tality of senile HAP was 29.2%. Conclusion There are high antibiotic resistance and high mortality of HAP in se-nile patients. Measures should be taken to modify the risk factors.

Surveillance of bacterial resistance and study on ?-lactamases in cefoxitin-resistant Klebsiella pneumoniae isolates / 中国医师进修杂志

Objective To investigate the resistance and ?-lactamase of cefoxitin-resistant Klebsiella pneumoniae. Methods The minimal inhibitory concentrations were determined by standard agar dilution.Isoelectric focusing was used to measure the PI(s) of an isolate,s ?-lactamase,AmpC and ESBLs activity was confirmed by a three-dimensional extract method. Results The resistant rates of 40 strains were as follows: imipenem and meropenem 0.0%,cefepime 20.6%,cefotaxime(22.5%),ceftazidime 60.0%.The most isolates were demonstrated two or more ?-lactamase bands by IEF.Of 39 strains tested,ESBLs was detected in 31(70.5%) strains and AmpC-type?-lactamase in 16(41.0%) strains by three dimensional extract test. Conclusions These cefoxitin-resistant Klebsiella pneumoniae produced two or more ?-lactamases.It is imperative for clinical microbiology laboratories to detect and research ?-lactamases,especially AmpC enzyme.

Purification of EGF-TCS recombinant fusion protein and its targeting action on human tumor cells in vitro / 第三军医大学学报

Objective To express and purify EGF-TCS fusion protein and observe the targeted and selective killing effect on cancer cells of the fusion protein.Methods The recombinant expression plasmid PQE30/EGF-TCS was transformed to E.coli.M15 and the fusion protein(EGF-TCS) was expressed.Ni-NTA Agrose affinity chromatography was used to purify the protein,flow cytometry to detect EGFR expression rate in cancer cells(BEL-7402,MCF-7,BGC-823) and normal liver LO2 cells,and the killing test to verify selective killing ability of EGF-TCS;The cell apoptosis detection by flow cytometry and microscopic observation were used to confirm the selective killing ability of EGF-TCS.Results Recombinant expression plasmid PQE30/EGF-TCS was expressed in E.coli.M15 stably and effectively.The purity of EGF-TCS was over 95% by chromatography.EGFR expression rate was highest in hepatoma cells BEL-7402(72.33%) and lowest in normal liver LO2 cells(5.51%).The killing ability of recombinant protein was more effective to cancer cells(IC50 of BEL-7402,MCF-7 and BGC-823 was 11.4,22.47 and 12.53 ?g/ml respectively) and was weak to normal cells(IC50 53.19 ?g/ml).Conclusion The recombinant protein EGF-TCS that induces apoptosis of cancer cells was successfully constructed by gene engineering technology.

Effects of TAE and TACE on HIF-1?,MVD and PCNA in rabbit liver carcinoma / 中国普通外科杂志

Objective To investigate expression and clinical significance of HIF-1?、MVD and PCNA after TAE or TACE for rabbit liver carcinoma,and to identify the mechanism of hepatic carcinoma cell proliferation after the operation.Methods Thirty rabbits were implanted with liver VX2 tumors for 1 week.Then they were randomly divided into 3 groups.Control group had hepatic arterial infusion of 0.5 to 0.8 mL saline.TAE group was embolized with 0.5 to 0.8 ml UFLP.TACE group was embolized with 0.5 to 0.8 ml UFLP mixed with 2 mg THP.Two weeks after treatment,HIF-1?、MVD and PCNA expression was detected.Results Growth of the liver tumor was found in the 3 groups,and there were 15,11 and 3 new tumors found in the control group,TAE group and TACE group respectively.The expressions of HIF-1?and PCNA in control group were higher than those in TACE group,but significantly lower than those in TAE group(P

Resistance to Antibiotics of Clinical Isolates of Klebsiella pneumoniae Producing TEM-116 ?-Lactamases / 中华医院感染学杂志

OBJECTIVE To understand characteristics of TEM-116 ?-lactamases through comparative study on resistance to antibiotics of clinical isolates of Klebsiella pneumoniae producing TEM-116 ?-lactamases.METHODS K.pneumoniae susceptibility to ?-lactamases was determined by disk diffusion tests,and their isoelectric points(PI) were detected using analytic isoelectric focusing(IEF),and resistance to antibiotics of clinical isolates of K.pneumoniae producing TEM-116 and TEM-1?-lactamases was studied.RESULTS Both of K.pneumoniae producing TEM-116 ?-lactamases and producing TEM-1 ?-lactamases were 100% resistant to AMP,and highly resistant to the first and second generation cephalosporin,but greatly susceptible to FEP and IPM.There was greatly difference between resistance to AMC,TZP,AMK,and GEN of clinical isolates of K.pneumoniae producing TEM-116 ?-lactamases and that of K.pneumoniae producing TEM-116 ?-lactamases,the TEM-116 isolates were higher resistant than TEM-1 isolates.Analytic IEF results showed that PI of TEM-116 ?-lactamases was 5.4,and most strains of K.pneumoniae TEM-116 ?-lactamases displayed two electrophoresis bands or more,only one strain of them just displayed one band,resistant to majority of antibiotics.CONCLUSIONS The results show that K.pneumoniae producing TEM-116 ?-lactamases are more resistant to antibiotics than K.pneumoniae producing TEM-1 ?-lactamases,and indicate TEM-116 ?-lactamases work as ESBLs.