In this study, pEGFP-N1 was chosen as the reporter plasmid and transferred into Ctenopharyngodon idellus kidney (CIK) cells by electroporation, and the optimal electroporation conditions were determined by testing the transfection efficiency with different voltages, pulse times, plasmid amounts, and numbers of shocks. The results showed that the maximum electroporation efficiency was achieved under the following conditions in a 0.2 cm electroporation cuvette containing CIK cells (1.5 x 10(7)/mL, 200 microl): electric voltage 200 V, pulse time 45 ms, plasmid 30 microg, and one electric shock. The total genomic RNA of grass carp reovirus (GCRV) was extracted in this experiment and reversely transcribed into cDNA, which was used to amplify the gene segment of GCRV non-structural protein NS26 using designed specific primers. The PCR product was recombined into pEGFP-N1 vector. The fusion protein EGFP-NS26 was successfully and efficiently expressed in the CIK cells by electroporation, which was confirmed by both fluorescent imaging and Western blot analysis. This experiment laid a foundation for further functional studies of the non-structural protein NS26 of GCRV.
Animals ; Cell Line ; Cyprinidae ; Electroporation ; Fish Diseases ; virology ; Gene Expression ; Kidney ; virology ; Reoviridae ; genetics ; physiology ; Reoviridae Infections ; veterinary ; virology ; Viral Nonstructural Proteins ; genetics ; metabolism

Objective To compare the effect of transthoracic contrast echocardiography (cTTE) with transesophageal contrast echocardiography (cTEE) for detecting right to left shunt (RLS) in patients with patent foramen ovale (PFO). Methods The prospective study was conducted in 29 consecutive patients with PFO who suffered from cryptogenic stroke and/or migraine. Contrast echocardiography was performed in all 29 patients. The cTTE was performed using transducer with second harmonic imaging modality (transmitting frequency 1.7 MHz, receiving frequency 3.4 MHz). The cTEE was performed using transducer with frequency 7 MHz. Ten milliliter saline solution of contrast were rapidly administrated through an antecubital vein. According to whether microbubble (MB) appearing in left atrium after complete opaciifcation of right atrium within the ifrst 3 circles, the results were classiifed by a four-level semi-quantitative categorization:Level 1 (no PFO-RSL), no MB in left atrium; Level 2 (small PFO-RSL) 1-10 MBs; Level 3 (medium PFO-RSL) 10-30 MBs;Level 4 (large PFO-RSL)>30 MBs. Results The total detection rate of PFO-RSL was signiifcant different between cTTE and cTEE (86.2%vs 55.2%,χ2=5.711, P=0.017). In cTTE there were 4 cases at level 1, 1 case at level 2, 5 cases at level 3 and 19 cases at level 4. In cTEE there were 13 cases at leverl 1, 2 cases at level 2, 6 cases at level 3 and 7 cases at level 4. The comparison of semi-quantitative grading derived from cTTE and cTEE was also signiifcant different (Wilcoxon signed ranks test showed Z=-3.789, P=0.000). Conclusions The efifciency in detection of PFO-RLS with cTTE was better than with cTEE. Compared with cTEE, cTTE was easier in practice and brought less discomfort and complications to patients.

Objective To evaluate the curative effect on chdecytolithiasis with Chinese traditional patent formulation Qingdanyin and ESWL. Methods From January 2000 to January 2003,332 patients with gallstones were treated with Qingdanyin and ESWL, compared with 141 patients with routine ESWL. Results The 2-week clean rate(36.14%),one month clean rate(58. 58%),3-month clean rate(68. 98%) ,6-month clean rate(86.75%) of gallstones in the Qingdanyin and ESWL group were much higher than those in routine ESWL patients(14. 92%, 21.02%,35.93%,47.11%,respectively)(P

The patients with dedifferentiated thyroid carcinoma were induced by all-trans retinoic acid (ATRA). The treatment was effective in 5 patients (5/9,55.6%), and 5 patients were benefited from following 131 I treatment(5/8, 62.5% ). Three patients(3/12,25.0% ) quitted because of neurological side-effects. It is efficacious to apply ATRA to induce dedifferentiated thyroid carcinoma and improve radio-iodine uptake and acquire benefit for patients, but clinic application of ATRA must be cautious.

Adult ; Brain Neoplasms ; pathology ; Glioblastoma ; pathology ; Humans ; Male ; Pineal Gland ; pathology

OBJECTIVETo study the effect of single administration of aqueous extracts from aconite on "dose-toxicity" relationship and "time-toxicity" relationship of mice hearts, through changes in electrocardiogram (ECG) and serum biochemical indexes.

METHODMice were grouped according to different drug doses and time points, and orally administered with water extracts from aconite for once to observe the changes of mice ECG before and after the administration, calculate visceral indexes heart, liver and kidney, and detect levels of CK, LDH, BNP and CTn-I in serum.

RESULTAccording to the "time-toxicity" relationship study, at 5 min after oral administration with aqueous extracts from aconite in mice, the heart rate of mice began rising, reached peak at 60 min and then slowly reduced; QRS, R amplitude, T duration and amplitude and QT interval declined at 5 min, reduced to the bottom at 60 min and then gradually elevated. The levels of CK, LDH, BNP and CTn-I in serum elevated at 5 min and reached the peak at 60 min, with no significant change in ratios of organs to body at different time points. On the basis of the "dose-toxicity" relationship, with the increase in single dose of aqueous extracts from aconite, the heart rate of mice. QRS, T duration and amplitude and QT interval declined gradually, and levels of CK, LDH, BNP and CTn-I in serum slowly elevated, with a certain dose dependence and no significant change in ratios of organs to body in mice.

CONCLUSIONSingle oral administration of different doses of aqueous extracts from aconite could cause different degrees of heart injury at different time points, with a certain dose dependence. Its peak time of toxicity is at 60 min after the administration of aqueous extracts from aconite.

Aconitum ; adverse effects ; chemistry ; Animals ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; toxicity ; Female ; Heart ; drug effects ; physiology ; Heart Rate ; drug effects ; Kidney ; drug effects ; Liver ; drug effects ; Male ; Mice

Objective To investigate the treatment of atypical hemolytic uremic syndrome (aHUS) in their acute phase and relate the prognostic factors.Methods Twenty-eight patients with aHUS treated in Department of Nephrology,Beijing Children's Hospital Affiliated to Capital Medical University from January 2011 to March 2015 were collected,the outcomes of the treatment were summarized.Results All the patients were given plasma exchange (PE)treatment,21 patients with hemodialysis (HD) treatment;13 patients with H factor (FH) antibody were further treated with corticosteroid therapy,and 1 patient was treated with the combination of mycophenolate mofetil.Evaluation of patients' indices were conducted at month 3.Out of the 28 patients,there were 13 patients with complete remission;of the 14 patients showed improvement,6 patients were complicated with hypertension and 4 patients with renal dysfunction.One patient,with ongoing hemodialysis,did not show restoration of renal function.There was no death in children.Statistical analysis showed that the acute phase serum creatinine levels were (430.0 ± 124.7) μmol/L and (265.6 ± 90.8)μmol/L for the group with remaining kidney injury and the group with complete remission,respectively,the difference was statistically significant(P ＜0.01).The levels of FH for the 2 groups were (206.5 ± 134.9) mg/L and(415.4-±214.1) mg/L,and the difference was statistically significant (P ＜ 0.01) as well.Statistical analysis showed that differences were no statistically significant between the 2 groups of patients on their age,platelets,complement C3,complement C4,hemoglobin and lactate dehydrogenase (all P ＞ 0.05).For the group with positive antibody,its average serum creatinine was (346.4 ± 112.5) μmol/L.For the group with negative antibody,its average serum creatinine was (297.0 ± 89.3) μmol/L.The FH for group with positive antibody and group with negative antibody were (307.4 ±187.3) mg/L and (379.8 ± 203.5) mg/L,respectively.The differences were not statistically significant (all P ＞0.05).Conclusions PE is an effective treatment for aHUS at acute phase,and corticosteroid and immunosuppressive therapy are effective for patients with positive anti-FH antibody.For patients with severe kidney injury at acute phase,the possibility of long-term kidney damage is high.Low concentration of serum FH indicates a poor prognosis.

Objective To analyze the clinical features of pediatric sepsis with initial symptoms of autoimmune hemolytic anemia(AIHA) and acute renal failure.Methods Ten cases of pediatric sepsis with initial symptoms of AIHA and acute renal failure were selected from January 2000 to January 2014 in Department of Nephrology,Beijing Children's Hospital Affiliated to Capital University of Medical Science.The clinical data were retrospectively analyzed and long-time follow-up was taken.Results There were 8 boys and 2 girls of the 10 cases.The ages ranged from 8 months to 12.8 years.There were 8 children less than 6 years old having a prevalence of 80％.There were sudden dark urine and progressive anemia in all the cases,jaundice in 9 cases,abdominal pain and vomiting in 6 cases.Urine volume decreased in 7 cases and anuria in 3 cases.Four cases showed disorder of consciousness and 2 developed shock.Respiratory failure and disseminated intravascular coagulation happened in 1 case respectively.In terms of pathogens,2 cases were infected by mycoplasma and 1 case was infected by hemolytic streptococcus.The rest of the pathogen was unknown.The Coomb's test was 3 +-4 + positive.Serum urea nitrogen and creatinine gradually increased.High level total serum bilirubin and unconjugated bilirubin were shown in 8 cases and low level complement in 7 cases.Bone marrow films showed hyperplasia in all the cases.Renal biopsy was taken in 3 cases and no microthrombus was found.Corticosteroid was used in all 10 cases for 6 months.Blood purification was taken in 8 cases.Respiratory support was used in 4 cases.Nine recovered and 1 case gave up treatment.Further follow-up was taken for 2.2-8.1 years.Eight cases had normal urine and blood routine test and renal function.Relapse happened in 1 girl 1 year later.One case lost follow-up.Conclusions Once sudden hemoglobinuria,anemia and progressively decreased urine output occur in sepsis cases,the clinicians should consider AIHA and acute renal failure.Based on active infection control,the early use of steroids and blood purification treatment can improve the prognosis.

Objective To analyze the clinical characteristics,prognosis and the risk factors in 48 children with lupus nephritis combined with renal insufficiency.Methods The clinical data from 48 pediatric lupus nephritis with renal insufficiency from January 2000 to January 2013 were retrospectively analyzed.Results Forty-eight cases showed renal insufficiency among 168 children with lupus nephritis,and the incidence rate was 28.6％.The age ranged from 5.8 to 16.2 years.The male to female ratio was 1.0 ∶ 2.2.Among 48 children,43 cases had hematuria,41 cases had heavy proteinuria,25 cases had anaemia and 23 cases had persistent hypertension.Totally 20 cases underwent renal biopsy,and among them,15 cases(75.0％) were diagnosed as diffuse proliferative lupus nephritis(class Ⅳ).The histological activity index was ≥7 in 13 cases and the histological chronicity index was ≥4 in 3 cases.Corticosteroid and/or cytotoxic drugs were used in all of 48 cases.Thirty-five cases had normal urine and renal function,5 cases had stable renal function with persistent proteinuria,4 cases developed into chronic renal failure and 4 cases died.Persistent hypertension (x2 =4.274,P =0.039),the time of starting therapy (x2 =28.830,P =0.000),and histologic chronicity index(P =0.008 8 by Fisher's exact probability test)were the prognostic factors.Conclusions Among pediatric lupus nephritis with renal insufficiency,class Ⅳ (diffuse proliferative lupus nephritis) is the most frequent finding.Persistent hypertension,the time of starting therapy,and histologic chronicity index are the prognostic factors.The outcomes of lupus nephritis with insufficiency can be improved by adequate and appropriate treatment.

Objective To explore the clinical features of invasive fungal infection in 27 children treated at nephrology department between 1999 and 2014.Methods Twenty-seven cases of invasive fungal infection at nephrology department were analyzed retrospectively.Results Candida urinary infection was found in 9 cases,pneumocystis carinii pneumonia were found in 8 cases,invasive pulmonary aspergillosis and urinary aspergillosis found in 4 cases and 1 case respectively,cryptococcus neoformans meningitis and pulmonary candidiasis found respectively in 2 cases,and pulmonary filamentous fungal disease was found in 1 case.These 27 cases showed different features of illness:10 primary nephrotic syndrome,7 secondary nephrotic syndrome,6 malformation of urinary development and 4 preterm birth ＜ 32 weeks gestational age with low birth weight.All of the cases had the history of taking antibiotics.Seventeen cases of them had used corticosteroid and cytotoxic drugs in a long-term.Sixteen cases had experienced invasive procedures.All children had fever in varying degrees,14 cases showed gasp,7 cases had progressive hypoxia and respiratory failure,and 5 cases developed into multiple organs failure.Chest computed tomographic (CT) imaging data showed diffuse ground-glass opacity with mosaic sign of pneumocystis carinii pneumonia(8 cases).The features of pulmonary aspergillosis included multiple nodules and cavity in bilateral lungs (4 cases).The radiologic findings showed patching lesions with indistinct edge and uncertained density of bilateral middle-lower lung fields with pulmonary candidiasis (2 cases).Full dose of antifungal drug was given to 23 cases of them,16 cases recovered completely,3 got better,4 cases died.Four cases gave up full dose antifungal therapy.Conclusions Long-term use of corticosteroid and cytotoxic drugs,use of broad-spectrum antibiotics,invasive therapies,such as indwelling centralvenous catheters,endotracheal intubation with mechanical ventilation,and preterm birth with low birth weight were risk factors in predicting invasive fungal infection.Chest CT findings were different among these cases.Even though not specific,the relatively differences were helpful to the differential diagnosis of these diseases.