Objective: To explore the false-negative possibility in genetic test of congenital long QT syndrome (LQTS) by next-generation sequencing (NGS). Methods: A total of 28 genomic DNA samples were collected from 4 laboratories including 2 commercial medical laboratories using HiSeq2000 platform as Lab1,n=6 and Lab2,n=8; 1 commercial research service laboratory using Ion-torrent platform as Lab3,n=8 and 1 academic laboratory using HiSeq2000 platform as Lab 4,n=6. Sequencing coverage in the exons of protein-coding region in 3 main LQTS pathogenic genes as KCNQ1, KCNH2, SCN5A and possible pathogenic variants were quantitatively analyzed. Results: In Lab1, Lab 2 and Lab 4 with HiSeq2000 platform, above 98% protein coding regions in 3 pathogenic genes were covered with>10-fold reads and 90%-95% were covered with>30-fold reads. In 2 commercial medical laboratories, 3.63% and 9.84% protein coding regions of KCNQ1 gene in 14 samples were covered with<10-fold reads and with<30-fold reads; lower than 10-fold covering region was focused in the 1st exon including about 2% known or likely pathogenic variants. In 2 commercial medical laboratories, 2.64% and 15.76% protein coding regions of KCNH2 gene in 14 samples were covered with<10-fold reads and with<30-fold reads; low covering region was located in multiple exons. For the data from Lab 1, as high as 28.56% protein coding regions of KCNH2 gene were covered with<30-fold reads including 113 (19.79%) known or likely pathogenic variants. SCN5A gene had the best coverage of protein coding region, with no<10-fold reads in all 4 Labs and no<30-fold reads in 2 commercial medical laboratories. Conclusion: Currently, NGS has low coverage region in both KCNQ1 and KCNH2 genes, pathogenic variants could be missed and false-negative possibility should be highly alert.

OBJECTIVETo investigate the expression of zinc ribbon domain-containing1 (ZNRD1) in human renal cell carcinoma and normal kidney tissues.

METHODSThe expression of ZNRD1 protein was examined by immunohistochemical staining in 71 renal cell carcinomas and 24 samples of normal kidney tissue. The correlation between the expressions of ZNRD1 protein and clinicopathologic features was analyzed. The expression of ZNRD1 mRNA and ZNRD1 protein was detected by quantitative reverse transcriptase-polymerase chain reaction (PT-PCR) and Western blot in 20 renal cell carcinomas and corresponding adjacent non-cancerous tissues.

RESULTSZNRD1 protein was detected mostly in the cell nuclei by immunohistochemistry. The positive expression rate of ZNRD1 protein was 91.7% (22/24) in renal cell carcinomas and 20.8% (5/24) in the normal kidney tissues, with a statistically significant difference between cancer and normal kidney tissue (P < 0.01). However, no significant correlation was observed between ZNRD1 protein expression level and clinicopathologic features (P > 0.05). ZNRD1 mRNA expression level was significantly higher in renal cell carcinomas (0.6186) than that in the normal kidney tissues (0.4273) assessed by RT-PCR (P < 0.01). The expression level of ZNRD1 protein by Western blot was 0.5623 in renal cell carcinomas, significantly higher than that in normal kidney tissues (0.3885, P < 0.01).

CONCLUSIONZNRD1 gene and ZNRD1 protein may play an important role in the carcinogenesis of renal cell carcinoma. Further investigation is still needed.

Adult ; Aged ; Aged, 80 and over ; Blotting, Western ; Carcinoma, Renal Cell ; metabolism ; pathology ; DNA-Binding Proteins ; biosynthesis ; genetics ; Female ; Humans ; Immunohistochemistry ; Kidney ; metabolism ; Kidney Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo constitute a model of B immunoblastic lymphomas in the Hu-PBL-SCID mice.

METHODSThe SCID mice were reconstituted by intraperitoneal injection (i.p.) of 5 x 10(7) human lymphocytes from Epstein-Barr virus (EBV) seronegative individuals. After one week, the SCID mice were inoculated with EBV by i.p. injection, and subjected to the investigation of whether there was any tumor in the abdomen of such SCID mice four weeks later. The characteristics of the found tumor was observed by the methods of Hematoxylin-eosin (HE) stain, immunohistochemical staining and polymerase chain reaction (PCR).

RESULTSCompared with the control groups, all the EBV-infected Hu-PBL-SCID mice had abdominal solid tumors [(32 +/- 12.5) mm3] developed, often located in the liver. HE staining and immunohistochemical staining showed the tumors were human B cell lymphomas. EBV DNA could be detected in the tumors by the PCR.

CONCLUSIONSThe model of B immunoblastic lymphomas in the Hu-PBL-SCID mice is successfully constituted, and may well be useful to the human tumor immunological study.

Animals ; Disease Models, Animal ; Herpesvirus 4, Human ; physiology ; Humans ; Lymphoma, Large-Cell, Immunoblastic ; Mice ; Mice, SCID

Objective To develop an ultra-performance liquid chromatography/quadrupole-time of flight mass spectrometry(UPLC-QTOF-MS)method for the determination of an oxadiazole-2-oxide heterocyclic compound F-2015-14.Methods Mouse plasma and liver homogenate specimens were extracted with ethyl acetate and chromatographed on a Waters CORTECS column(C18,1.6μm,2.1 mm×150 mm)by using a mobile phase of 10％acetonitrile-0.1％formic acid with by a volume fractionation by gradient elution.Then,UPLC-QTOF-MS was performed to determine F-2015-14 in mouse plasma and liver homogenate speci-mens.Results The linearity of F-2015-14 in plasma ranged from 12.5 to 250 μg/mL with a correlation coefficient of 0.990 and a detection limit of 8.8 μg/mL.F-2015-14 in liver homogenates ranged from 12.5 to 250 μg/mL.The linearity was good with a cor-relation coefficient of 0.992 and a limit of detection of 5.6 μg/mL.If the concentration of plasma and liver homogenate specimens was 12.5 μg/mL,the accuracy and the matrix effect were 80％to 120％,and the inter-day and intra-day precision was within 20％.If the concentrations of plasma and liver homogenate specimens were 100 μg/mL and 200 μg/mL,the accuracy and the ma-trix effect were 85％to 115％,and the inter-day and intra-day precision was within 15％.Conclusion The UPLC-QTOF-MS es-tablished in this study has a high sensitivity and good reproducibility for the determination of F-2015-14,which provides bases for the development of novel anti-schistosomiasis drugs.

Animals ; Cyclooxygenase 2 ; metabolism ; Lipopolysaccharides ; Liver Failure ; metabolism ; Male ; Mice ; Mice, Inbred BALB C

In the present study, the safety of Haemophilus influenza type b conjugate vaccines inoculated in the upper arm deltoid and vastus lateralis muscle was evaluated. 680 infants aged 2-5 months and 6-12 months were selected to be the research subjects in whom the Hib conjugate vaccines were inoculated by injection in the upper arm deltoid and vastus lateralis muscle, respectively. The safety analysis indicated that there were no statistic differences in the incidence rates of adverse reactions when the Hib conjugate vaccines were inoculated at different sites. So we concluded that the safety of inoculation injection of Hib conjugate vaccines in vastus lateralis muscle was the same as that inoculated in the upper arm deltoid.
Bacterial Capsules ; China ; Haemophilus Vaccines ; administration & dosage ; adverse effects ; Humans ; Incidence ; Infant

Objective To evaluate the safety and efficacy of the CARTO3-based total three-dimensional(T3D,total three-dimensional)zero X-ray mapping technique in radiofrequency catheter ablation of elderly patients with atrial fibrillation. Methods A total of 60 patients diagnosed with paroxysmal atrial fibrillation underwent radiofrequency catheter ablation at the Beijing Anzhen Hospital Arrhythmia Center from December 2015 to April 2017 were included. All patients were randomly divided into the study group(30 cases) and the control group(30 cases). T3D technique was utilized in the study group, and patients in the control group received conventional AF ablation. The procedure parameters success rate of circumferential pulmonary vein isolation(CPVI), rates of atrial fibrillation recurrence complication were compared between the two groups. Results All the 60 patients with paroxysmal atrial fibrillation had successful atrial fibrillation ablation and finished follow-up. Compared with the control group, the time of atrium three-dimensional reconstruction in the study group was longer [study group vs control group:(57.7±11.0)min vs.(10.4±3.5)min,P<0.001)];X-ray exposure time was significantly shorter in the study group[study group vs control group:0 min vs.(15.73±3.91)min,(P<0.001)]. The diff erence in circumferential pulmonary vein ablation time between the two groups was not of statistical significance[study group vs. control group:(49.9 ± 11.3)min vs.(51.1 ± 12.6)min,P=0.699].CPVI was successful in all patients in both groups. There was no signifi cant diff erence in the early and late recurrence rate and the incidence of complications between the two groups(P>0.05). Conclusions The application of T3D technique in radiofrequency ablation for elderly patients with paroxysmal atrial fi brillation is safe and eff ective, which can reduce the time of X-ray exposure and has important clinical value.

OBJECTIVE: To evaluate the storage stability of metabolites from actinomycetes Streptomyces nigrogriseolus XD 2-7 and the mollcuscicidal activity against Oncomelania hupensis in the laboratory, and to preliminarily explore the mechanisms of the molluscicidal activity. METHODS: The fermentation supernatant of S. nigrogriseolus XD 2-7 was prepared and stored at -20, 4 °C and 28 °C without light for 10 d; then, the molluscicidal effect was tested against O. hupensis following immersion for 72 h. The fermentation supernatant was boiled in a 100 °C water bath for 30 min and recovered to room temperature, and then the molluscicidal effect was tested against O. hupensis following immersion for 72 h. The pH values of the fermentation supernatant were adjusted to 4.0, 6.0 and 9.0 with concentrated hydrochloric acid and sodium hydroxide, and the fermentation supernatant was stilled at room temperature for 12 h, with its pH adjusted to 7.0; then, the molluscicidal effect was tested against O. hupensis following immersion for 72 h. The fermentation product of S. nigrogriseolus XD 2-7was isolated and purified four times with macroporous resin, silica gel and octadecylsilane bonded silica gel. The final products were prepared into solutions at concentrations of 10.00, 5.00, 2.50, 1.25 mg/L and 0.63 mg/L, and the molluscicidal effect of the final productswas tested against O. hupensis following immersion for 72 h, while dechlorination water served as blank controls, and 0.10 mg/L niclosamide served as positive control. The adenosine triphosphate (ATP) and adenosine diphosphate (ADP) levels were measured in in O. hupensis soft tissues using high performance liquid chromatography (HPLC) following exposure to the final purified fermentation products of S. nigrogriseolus XD 2-7. RESULTS: After the fermentation supernatant of S. nigrogriseolus XD 2-7 was placed at -20, 4 °C and 28 °C without light for 10 d, immersion in the stock solution and solutions at 10- and 50-fold dilutions for 72 h resulted in a 100% (30/30) O. hupensis mortality. Following boiling at 100 °C for 30 min, immersion in the stock solution and solutions at 10- and 50-fold dilutions for 72 h resulted in a 100.00% (30/30) O. hupensis mortality. Following storage at pH values of 4.0 and 6.0 for 12 h, immersion in the fermentation supernatant of S. nigrogriseolus XD 2-7 for 72 h resulted in a 100.00% (30/30) O. hupensis mortality, and following storage at a pH value of 9.0 for 12 h, immersion in the fermentation supernatant of S. nigrogriseolus XD 2-7 for 72 h resulted in a 33.33% (10/30) O. hupensis mortality (χ² = 30.000, P < 0.05). The minimum concentration of the final purified fermentation products of S. nigrogriseolus XD 2-7 was 1.25 mg/L for achieving a 100% (30/30) O. hupensis mortality. The ATP level was significantly lower in O. hupensis soft tissues exposed to 0.10 mg/L and 1.00 mg/L of the final purified fermentation products of S. nigrogriseolus XD 2-7 than in controls (F = 7.274, P < 0.05), while no significant difference was detected in the ADP level between the treatment group and controls (F = 2.485, P > 0.05). CONCLUSIONS The active mollcuscicidal ingredients of the S. nigrogriseolus XD 2-7 metabolites are maintained stably at -20, 4 °C and 28 °C for 10 d, and are heat and acid resistant but not alkali resistant. The metabolites from S. nigrogriseolus XD 2-7 may cause energy metabolism disorders in O. hupensis, leading to O. hupensis death.
Adenosine Diphosphate/pharmacology* ; Adenosine Triphosphate ; Animals ; Molluscacides/pharmacology* ; Silica Gel/pharmacology* ; Snails ; Streptomyces ; Water

BACKGROUNDData on the epidemiology of hypertension in Chinese non-dialysis chronic kidney disease (CKD) patients are limited. The aim of the present study was to investigate the prevalence, awareness, treatment, and control of hypertension in the non-dialysis CKD patients through a nationwide, multicenter study in China.

METHODSThe survey was performed in 61 tertiary hospitals in 31 provinces, municipalities, and autonomous regions in China (except Hong Kong, Macao, and Taiwan). Trained physicians collected demographic and clinical data and measured blood pressure (BP) using a standardized protocol. Hypertension was defined as systolic BP ≥ 140 mmHg and/or diastolic BP ≥ 90 mmHg, and/or use of antihypertensive medications. BP < 140/90 mmHg and < 130/80 mmHg were used as the 2 thresholds of hypertension control. In multivariate logistic regression with adjustment for sex and age, we analyzed the association between CKD stages and uncontrolled hypertension in non-dialysis CKD patients.

RESULTSThe analysis included 8927 non-dialysis CKD patients. The prevalence, awareness, and treatment of hypertension in non-dialysis CKD patients were 67.3%, 85.8%, and 81.0%, respectively. Of hypertensive CKD patients, 33.1% and 14.1% had controlled BP to < 140/90 mmHg and < 130/80 mmHg, respectively. With successive CKD stages, the prevalence of hypertension in non-dialysis CKD patients increased, but the control of hypertension decreased (P < 0.001). When the threshold of BP < 130/80 mmHg was considered, the risk of uncontrolled hypertension in CKD 2, 3a, 3b, 4, and 5 stages increased 1.3, 1.4, 1.4, 2.5, and 4.0 times compared with CKD 1 stage, respectively (P < 0.05). Using the threshold of < 140/90 mmHg, the risk of uncontrolled hypertension increased in advanced stages (P < 0.05).

CONCLUSIONSThe prevalence of hypertension Chinese non-dialysis CKD patients was high, and the hypertension control was suboptimal. With successive CKD stages, the risk of uncontrolled hypertension increased.

Adult ; Aged ; Awareness ; Female ; Humans ; Hypertension ; complications ; epidemiology ; therapy ; Male ; Middle Aged ; Prevalence ; Renal Insufficiency, Chronic ; complications

OBJECTIVE: Preliminary assessment of rabies virus neutralizing activity, safety and immunogenicity of a recombinant human rabies antibody (NM57) compared with human rabies immunoglobulin (HRIG) in Chinese healthy adults. METHODS: Subjects were randomly (1:1:1) allocated to Groups A (20 IU/kg NM57), B (40 IU/kg NM57), or C (20 IU/kg HRIG). One injection was given on the day of enrollment. Blood samples were collected on days -7 to 0 (pre-injection), 3, 7, 14, 28, and 42. Adverse events (AEs) and serious AEs (SAEs) were recorded over a period of 42 days after injection. RESULTS: All 60 subjects developed detectable rabies virus neutralizing antibodies (RVNAs) (> 0.05 IU/mL) on days 3, 7, 14, 28, and 42. The RVNA levels peaked on day 3 in all three groups, with a geometric mean concentration (GMC) of 0.2139 IU/mL in Group A, 0.3660 IU/mL in Group B, and 0.1994 IU/mL in Group C. At each follow-up point, the GMC in Group B was significantly higher than that in Groups A and C. The areas under the antibody concentration curve over 0-14 days and 0-42 days in Group B were significantly larger than those in Groups A and C. Fifteen AEs were reported. Except for one grade 2 myalgia in Group C, the other 14 were all grade 1. No SAEs were observed. CONCLUSION The rabies virus neutralizing activity of 40 IU/kg NM57 was superior to that of 20 IU/kg NM57 and 20 IU/kg HRIG, and the rabies virus neutralizing activity of 20 IU/kg NM57 and 20 IU/kg HRIG were similar. Safety was comparable between NM57 and HRIG.
Adult ; Antibodies, Neutralizing ; Antibodies, Viral ; Data Collection ; Humans ; Rabies/prevention & control* ; Rabies Vaccines/adverse effects* ; Rabies virus/genetics*