Publications by authors named "Bihong Huang"

22 Publications

  • Page 1 of 1

Development of a recombinase polymerase amplification fluorescence assay to detect feline coronavirus.

Mol Cell Probes 2020 12 22;54:101669. Epub 2020 Oct 22.

Guangdong Laboratory Animals Monitoring Institute and Guangdong Provincial Key Laboratory of Laboratory Animals, Guangzhou, 510633, China. Electronic address:

Feline coronavirus (FCoV) is classified into two pathotypes: the avirulent feline enteric coronavirus (FECV), and the virulent feline infectious peritonitis virus (FIPV). Rapid pathogen detection, which is efficient and convenient, is the best approach for early confirmatory diagnosis. In this study, we first developed and evaluated a rapid recombinase polymerase amplification (RPA) detection method for FCoV that can detect FCoV within 15 min at 39 °C. The detection limit of that assay was 233 copies/μL DNA molecules per reaction. The specificity was high: it did not cross-react with canine distemper virus (CDV), canine coronavirus (CCoV), canine adenovirus (CAV), feline calicivirus (FCV), feline herpesvirus (FHV), or feline parvovirus (FPV). This assay was evaluated using 42 clinical samples (30 diarrhea samples and 12 ascites samples). The coincidence rate between FCoV-RPA and RT-qPCR for detection in clinical samples was 95.2%. In summary, FCoV-RPA analysis provides an efficient, rapid, and sensitive detection method for FCoV.
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http://dx.doi.org/10.1016/j.mcp.2020.101669DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7581357PMC
December 2020

Application of recombinase polymerase amplification method for rapid detection of infectious laryngotracheitis virus.

Mol Cell Probes 2020 12 3;54:101646. Epub 2020 Aug 3.

Guangdong Laboratory Animals Monitoring Institute and Guangdong Provincial Key Laboratory of Laboratory Animals, Guangzhou, 510633, China. Electronic address:

Infectious laryngotracheitis is a significant respiratory disease of chickens that causes huge economic losses due to high morbidity and mortality and reduced egg production. A real-time recombinase polymerase amplification (RPA) assay was developed to accurately detect ILTV. The specific probe and primer sets were carefully designed and screened. The real-time RPA assay was carried out at 39 °C for 30 min, and results were obtained within 15 min. The results of the specificity assay showed no fluorescence signals with other avian-related viruses. The sensitivity of the assay was 1 × 10 copies/μL. The low CV value showed that the assay was reproducible. A total of 115 clinical samples were tested using the real-time RPA assay and the real-time PCR assay in parallel; the coincidence rates of the two detection methods were 100%. The results indicated that the real-time RPA assay is a specific, sensitive, rapid, and useful tool for epidemiological studies and clinical diagnosis, especially in the field and in resource-poor areas.
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http://dx.doi.org/10.1016/j.mcp.2020.101646DOI Listing
December 2020

Risk factors for decreased upper-limb muscle strength and its impact on survival in maintenance hemodialysis patients.

Int Urol Nephrol 2020 Jun 4;52(6):1143-1153. Epub 2020 May 4.

Division of Nephrology, National Clinical Research Center for Aging and Medicine, Huashan Hospital, Shanghai Medical College, Fudan University, No. 12 Wulumuqi Road, Shanghai, 200040, China.

Purpose: Protein-energy wasting, characterized by decreased muscle mass, is one of the strongest predictors of mortality in patients on maintenance hemodialysis (MHD). As people get older, their muscle strength usually declines faster than muscle mass. However, the association between lower-limb muscle strength and all-cause mortality remains unclear. We aimed to evaluate risk factors for decreased upper-limb muscle strength in MHD patients and its impact on patient survival.

Methods: The cross-sectional part of the study included 174 MHD patients. Subsequently, they were followed up for 52 weeks. Biceps muscle strength, anthropometry, body composition, dietary intake, daily steps, and biochemical indicators of malnutrition and inflammation were evaluated. Risk factors for muscle weakness were screened by multiple linear regression analysis, and patient survival was analyzed by Kaplan-Merier and Cox multivariate analysis.

Results: The 174 MHD patients (93 men; 63.05 ± 12.29 years) were classified as a young (< 65 years, n = 97) group and an elderly group (≥ 65 years, n = 77). Gender, daily steps, muscle mass, 25(OH)D level and IL-6 in young group, and muscle mass, 25(OH)D, daily steps, and NT-proBNP in elderly group were associated with the decreased biceps muscle strength. The survival rate in high muscle strength group was significantly higher than that in low muscle strength group (P = 0.002). The association between low muscle strength and high mortality risk remained strong in the fully adjusted model.

Conclusion: Risk factors of muscle weakness were different between young and elderly MHD patients. There was a strong correlation between strong biceps muscle strength and high patient survival.
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http://dx.doi.org/10.1007/s11255-020-02468-0DOI Listing
June 2020

Development of a real-time reverse transcription recombinase polymerase amplification assay for rapid detection of spring viremia of carp virus.

Mol Cell Probes 2020 04 19;50:101494. Epub 2019 Dec 19.

Key Laboratory of Fishery Drug Development, Ministry of Agriculture, Key Laboratory of Aquatic Animal Immune Technology, Guangdong Province, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, 510380, China. Electronic address:

Spring viremia of carp virus (SVCV) is a significant pathogenic agent that can cause large-scale outbreaks of spring viremia of carp (SVC) in many types of fish and bring huge economic losses to the aquaculture industry. A simple and convenient detection method is imperative for SVCV diagnosis. In this study, the real-time reverse transcription recombinase polymerase amplification (RT-RPA) assay was developed and validated. Primers and probe targeting the conserved region of M gene were designed and applied to the real-time RT-RPA assay that performed at 39 °C for 20 min. The specificity analysis showed that no cross-reaction with other pathogenic viruses of fish was found, indicating appropriate specificity of the assay. In vitro transcribed RNA standards were used to estimate the sensitivity of the assay and the detection limit was 10copies/reaction. To further evaluate the assay, 65 clinical samples were tested using both real-time RT-RPA assay and real-time RT-PCR method. The same detection results were observed, suggesting the potential application of real-time RT-RPA assay in clinical sample detection. This is the first report on RPA assay for SVCV detection and this new developed assay would be useful in both laboratory and in the field for diagnosis of SVCV.
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http://dx.doi.org/10.1016/j.mcp.2019.101494DOI Listing
April 2020

High-throughput Luminex xMAP assay for simultaneous detection of antibodies against rabbit hemorrhagic disease virus, Sendai virus and rabbit rotavirus.

Arch Virol 2019 Jun 15;164(6):1639-1646. Epub 2019 Apr 15.

Guangdong Laboratory Animal Monitoring Institute, Guangzhou, 510633, China.

Rabbits are widely used as models in biological research, and the pathogen status of rabbits used in studies can directly affect the results of experiments. Serological surveillance is the common monitoring method used in laboratory animals. A rapid, sensitive, and cost-effective high-throughput Luminex xMAP assay could be an attractive alternative to labor-intensive enzyme-linked immunosorbent assay (ELISA) methods. In this study, recombinant proteins from rabbit hemorrhagic disease virus and rabbit rotavirus and whole viral lysates of Sendai virus were used as coating antigens in an xMAP assay for the simultaneous detection of antibodies against these pathogens. The xMAP assay showed high specificity, with no cross-reaction with other pathogens. The coefficient of variation for intra-assay and inter-assay comparisons was less than 3% and 4%, respectively, indicating good repeatability and stability of the assay. The xMAP assay exhibited similar limits of detection for rabbit hemorrhagic virus and Sendai virus and was less sensitive for the detection of rabbit rotavirus when compared with commercial ELISA kits. A total of 52 clinical samples were tested simultaneously using both the xMAP assay and ELISA kits. The results obtained using these two methods were 100% coincident. In summary, the novel xMAP assay offers an alternative choice for rapid and sensitive high-throughput detection of antibodies in rabbit serum and can be used as a daily monitoring tool for laboratory animals.
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http://dx.doi.org/10.1007/s00705-019-04226-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7087182PMC
June 2019

Pharmacokinetics and pharmacodynamics of cinacalcet in patients with renal failure receiving hemodialysis and hemodiafiltration therapy
.

Int J Clin Pharmacol Ther 2019 Jun;57(6):284-289

Objective: Few studies have focused on the effects of dialysis on cinacalcet. In addition, there is no data available on hemodiafiltration (HDF) all over the world. Therefore, we studied the pharmacokinetics (PK) and pharmacodynamics (PD) of cinacalcet in patients undergoing hemodialysis (HD) or HDF to provide more guiding information on its use in these patients, especially in China.

Materials And Methods: In this open-label, single-dose, single-center study of 7 patients with renal failure who underwent dialysis, patients were randomly allocated to two groups consisting of 4 and 3 patients who received low-flux HD and HDF treatments, respectively. All participants underwent dialysis for 4 hours immediately after receiving single oral doses of a 25 mg cinacalcet tablet. High-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) and electrochemical luminescence (EI) were used to determine the cinacalcet plasma concentrations and intact parathyroid hormone (iPTH) serum levels, respectively.

Results: Peak concentration (C) and area under the curve (AUC) from time 0 to 24 hours (AUC) of the low-flux HD therapy group were 21.8 ± 18.6 ng/mL and 145.3 ± 91.8 ng×h/mL, respectively, which were similar to those of the HDF group (30.9 ± 7.9 ng/mL and 161.6 ± 26.5 ng×h/mL, respectively). iPTH concentrations of the HD therapy group decreased after cinacalcet administration and increased following its clearance. However, iPTH levels of subjects receiving HDF therapy did not change.

Conclusion: Compared with healthy Chinese subjects, patients with renal failure had a higher C and AUC, slightly prolonged time to C (t) after administration of the same dose of cinacalcet. On HD treatment day, variation trends of iPTH in Chinese patients and healthy subjects were similar and significantly different from that on the HDF treatment day. Considering the high protein binding rate of cinacalcet, this may lead to the great free-drug clearance during HDF treatment.
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http://dx.doi.org/10.5414/CP203260DOI Listing
June 2019

Point-of-care diagnostic assay for rapid detection of porcine deltacoronavirus using the recombinase polymerase amplification method.

Transbound Emerg Dis 2019 May 1;66(3):1324-1331. Epub 2019 Apr 1.

Guangdong Key Laboratory of Laboratory Animals, Guangdong Laboratory Animals Monitoring Institute, Guangzhou, China.

Porcine deltacoronavirus (PDCoV) has emerged and spread throughout the porcine industry in many countries over the last 6 years. PDCoV caused watery diarrhoea, vomiting and dehydration in newborn piglets. A sensitive diagnostic method would be beneficial to the prevention and control of PDCoV infection. Recombinase polymerase amplification (RPA) is an isothermal amplification method which has been widely used for virus detection. A probe-based reverse transcription RPA (RT-RPA) assay was developed for real-time detection of PDCoV. The amplification can be finished in 20 min and fluorescence monitoring was performed by a portable device. The lowest detection limit of the PDCoV RT-RPA assay was 100 copies of RNA molecules per reaction; moreover, the RT-RPA assay had no cross-reaction with other common swine viruses. The clinical performance of the RT-RPA assay was evaluated using 108 clinical samples (54 intestine specimens and 54 faecal swab specimens). The coincidence rate of the detection results for clinical samples between RT-RPA and RT-qPCR was 97.2%. In summary, the real-time RT-RPA assay offers a promising alternative to RT-qPCR for point-of-care detection of PDCoV.
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http://dx.doi.org/10.1111/tbed.13155DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7168525PMC
May 2019

Development of a SYBR green-based real-time RT-PCR assay for rapid detection of the emerging swine acute diarrhea syndrome coronavirus.

J Virol Methods 2019 03 26;265:66-70. Epub 2018 Dec 26.

Guangdong Key Laboratory of Laboratory Animals, Guangdong Laboratory Animals Monitoring Institute, Guangzhou, China. Electronic address:

Swine acute diarrhea syndrome coronavirus (SADS-CoV) is a novel coronavirus which was associated with severe diarrhea disease in pigs. SADS-CoV was first detected and identified as the causative agent of a devastating swine disease outbreak in southern China in 2017. Routine monitoring and early detection of the source of infection is therefore integral to the prevention and control of SADS-CoV infection. In this study, a SYBR green-based real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) technique was established for rapid detection and monitoring of this emerging virus. Specific primers were designed based on the conserved region within the M gene of the viral genome. The lowest detection limit of the RT-qPCR assay was 10 copies/μL. This assay was specific and had no cross-reaction with other 11 swine viruses. The positive rate of 84 clinical samples for the SYBR green-based RT-qPCR and the conventional RT-PCR was 73.81% (62/84) and 53.57% (45/84), respectively. These results demonstrated that the SYBR green-based RT-qPCR technique was an effectively diagnostic method with higher sensitivity than probe-based RT-qPCR and gel-based RT-PCR for detection and epidemiological investigations of SADS-CoV.
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http://dx.doi.org/10.1016/j.jviromet.2018.12.010DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7113735PMC
March 2019

Development and evaluation of a broadly reactive reverse transcription recombinase polymerase amplification assay for rapid detection of murine norovirus.

BMC Vet Res 2018 Dec 14;14(1):399. Epub 2018 Dec 14.

Guangdong Key Laboratory of Laboratory Animals, Guangdong Laboratory Animals Monitoring Institute, Guangzhou, China.

Background: Murine norovirus (MNV) is recognized as the most prevalent viral pathogen in captive mouse colonies. The rapid detection assay for MNV would be a useful tool for monitoring and preventing MNV infection. A recombinase polymerase amplification (RPA) assay was established in this study to provide a solution for rapid and sensitive detection of MNV.

Results: The detection limit of the RT-RPA assay for the detection of MNV was 1 × 10 copies of RNA molecules per reaction. The assay was specific since there was no cross-reaction with other common murine viruses. In addition, the broad reactivity of the RT-RPA assay was validated using the synthesized template carrying seven point mutations among several MNV strains. The MNV RT-RPA assay could detect as few as 1 × 10 copies of the mutant per reaction, suggesting the assay could be broadly reactive against a large diversity of MNV strains. Forty eight clinical samples including 16 gastric tissue specimens, 16 cecal tissue specimens and 16 fecal specimens were tested for the validation of the new developed RT-RPA assay. The detection results of RT-RPA and RT-qPCR for clinical samples were very similar, except that a gastric tissue sample which was positive by RT-qPCR, with a RNA titer of 27 copies, was negative by RT-RPA.

Conclusions: A broadly reactive RT-RPA assay was successfully established for MNV detection.
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http://dx.doi.org/10.1186/s12917-018-1736-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6295035PMC
December 2018

Development of a Conventional RT-PCR Assay for Rapid Detection of Porcine Deltacoronavirus with the Same Detection Limit as a SYBR Green-Based Real-Time RT-PCR Assay.

Biomed Res Int 2018 6;2018:5035139. Epub 2018 Nov 6.

Guangdong Key Laboratory of Laboratory Animals, Guangdong Laboratory Animals, Monitoring Institute, Guangzhou, China.

Porcine deltacoronavirus (PDCoV) is a newly discovered coronavirus, which belongs to the family Coronaviridae. It causes watery diarrhea, vomiting, and dehydration in newborn piglets. A sensitive RT-PCR method is urgently required to detect PDCoV infection. In this study, we developed and evaluated a conventional RT-PCR assay and a SYBR green-based real-time RT-PCR assay that targeted the PDCoV gene. Both assays are specific and have the same limit of detection at 2 × 10 copies of RNA molecules per reaction. Eighty-four clinical samples were subjected to both conventional RT-PCR and real-time RT-PCR, and the same positive rate (41.7%) was achieved, which was much higher than the positive rate (26.2%) using a previously described one-step RT-PCR technique. In summary, a conventional RT-PCR technique was successfully established for the detection of PDCoV with the same detection limit as a SYBR green-based real-time RT-PCR assay.
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http://dx.doi.org/10.1155/2018/5035139DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6247729PMC
April 2019

Application of ultrasonography in monitoring the complications of autologous arteriovenous fistula in hemodialysis patients.

Medicine (Baltimore) 2018 Nov;97(44):e12994

Department of Nursing, Huashan Hospital.

This study aims to evaluate the application of color Doppler ultrasound in monitoring the complications of autologous arteriovenous fistula in hemodialysis patients.Patients with maintenance hemodialysis who underwent autologous arteriovenous fistula were enrolled in this cross-sectional study. Ultrasound was used to detect fistula complications (stenosis and thrombosis), brachial artery diameter, and hemodynamic parameters. The ultrasound parameters were analyzed and screened to identify the most important indicator for monitoring complications.In all, 89 patients were included. Ultrasound showed 72 cases (80.90%) had normal fistula structure, and 17 cases (19.10%) had complications. The diameter, time-averaged mean velocity, flow volume, and diastolic peak velocity of brachial artery in complication group were significantly lower than those of noncomplication group (P < .05). The brachial artery pulsatility index and resistance index of complication group were significantly higher than those of noncomplication group (P < .05). There was no significant difference in peak flow velocity between complication and noncomplication group (P > .05). Indicators showed statistical significance were grouped based on quantiles. The incidence of complications was higher when the brachial artery diameter was ≤5.40 mm, or brachial artery flow was ≤460 mL/ min, or brachial artery pulsatility index was >1.04, or brachial artery resistance index was >0.60.Ultrasound monitoring of brachial artery diameter and hemodynamic parameters can help early detection of fistula complications. When the brachial artery diameter was ≤5.40 mm, or brachial artery flow was ≤460 mL/min, or brachial artery pulsatility index was >1.04, or brachial artery resistance index >0.60, stenosis or thrombosis should be checked to prevent fistula failure.
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http://dx.doi.org/10.1097/MD.0000000000012994DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6221746PMC
November 2018

The protective effect of different dialysis types on residual renal function in patients with maintenance hemodialysis: A systematic review and meta-analysis.

Medicine (Baltimore) 2018 Sep;97(37):e12325

Nursing Department, Huashan Hospital Affiliated to Fudan University, Shanghai, China.

Background: Residual renal function (RRF) is an important determinant of mortality and morbidity in patients undergoing hemodialysis. Different dialysis types may have different effects on RRF. We therefore conducted this meta-analysis to examine the RRF protective effect of different dialysis types for hemodialysis patients.

Methods: A systematic search was performed on PubMed, EMbase, Web of Science, Chinese Biomedical Literature Database, Wanfang database, and China National Knowledge Infrastructure for randomized controlled trials and cohort studies. Dialysis types included low-flux hemodialysis (LFHD), high-flux hemodialysis (HFHD), hemodiafiltration (HDF), and hemodialysis and hemoperfusion (HD+HP). The mean of endogenous creatinine clearance rate (CCR) and urea clearance rate (Curea), or urine volume was used to estimate RRF [95% confidence interval (95% CI), 6.05-16.80].

Results: There were 12 articles involving 1224 patients, including 11 random controlled trials and 1 cohort study. Meta-analysis showed that the RRF protective effect of HFHD [mean difference (MD) = 1.48, 95% CI (2.11 to 0.86), P < .01] and HD+HP [MD = 0.41, 95% CI (0.69 to 0.12), P = .005] was better than that of LFHD, and the RRF decline rate was the lowest in HFHD group [MD = 0.13, 95% CI (0.17 to 0.09), P < .01]. Descriptive analysis showed that HDF could better protect RRF when compared with LFHD. However, there was no consistency among other interventions when removing LFHD due to limited data.

Conclusion: For patients undergoing maintenance hemodialysis, the HFHD, HD+HP and HDF may better protect RRF, compared with LFHD.
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http://dx.doi.org/10.1097/MD.0000000000012325DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6156018PMC
September 2018

Rapid detection of three rabbit pathogens by use of the Luminex x-TAG assay.

BMC Vet Res 2018 Apr 7;14(1):127. Epub 2018 Apr 7.

Guangdong key laboratory of laboratory Animals, Guangzhou, China.

Background: Domestic rabbits especially New Zealand white rabbits play an important role in biological research. The disease surveillance and quality control are essential to guarantee the results of animal experiments performed on rabbits. Rabbit hemorrhagic disease virus, rabbit rotavirus and Sendai virus are the important pathogens that needed to be eliminated. Rapid and sensitive method focus on these three viruses should be established for routine monitoring. The Luminex x-TAG assay based on multiplex PCR and fluorescent microsphere is a fast developing technology applied in high throughput detection. Specific primers modified with oligonucleotide sequence/biotin were used to amplify target fragments. The conjugation between oligonucleotide sequence of the PCR products and the MagPlex-TAG microspheres was specific without any cross-reaction, and the hybridization products could be analyzed using the Luminex 200 analyzer instrument. Recombinant plasmids were constructed to estimate the detection limit of the viruses. Furthermore, 40 clinical samples were used to evaluate the efficiency of this multiplex PCR based Luminex x-TAG assay.

Results: According to the results, this new method showed high specificity and good stability. Assessed by the recombinant plasmids, the detection limit of three viruses was 100copies/μl. Among 40 clinical specimens, 18 specimens were found positive, which was completely concordant with the conventional PCR method.

Conclusions: The new developed Luminex x-TAG assay is an accurate, high throughput method for rapid detection of three important viruses of rabbits.
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http://dx.doi.org/10.1186/s12917-018-1438-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5889542PMC
April 2018

Comparative proteomic analysis of chief and oxyphil cell nodules in refractory uremic hyperparathyroidism by iTRAQ coupled LC-MS/MS.

J Proteomics 2018 05 9;179:42-52. Epub 2018 Mar 9.

Division of Nephrology, Huashan Hospital, Fudan University, Shanghai 200040, China. Electronic address:

SHPT is one of the most common complications of CKD-MBD. Recent studies indicate that oxyphil cell proliferation is related to SHPT progression, while not inhibited by current treatments. The aim of this study was to analyze the correlation between oxyphil cell and clinical indicators in SHPT, further explore the protein expression differences of oxyphil cell. Among 33 MHD patients, 84.8% patients have one or more oxyphil dominant glands and the overall oxyphil cells proportion was 39.5 ± 16.3%. Univariate correlation and multivariable linear regression model showed that oral calcitriol dosage and treatment duration were independently correlated to oxyphil cell ratio. Proteomic study showed that mitochondrial protein, protein synthesis, and cell cycle regulation were significantly altered in oxyphil cell nodules. DBP was downregulated in oxyphil nodules on protein level, which may contribute to calcitriol resistance by reducing vitamin D transport. Through KEGG and PPI network analysis, Wnt signaling, TGF-β, ubiquitin mediated proteolysis and cell cycle pathways were significantly enriched in oxyphil cell nodules. Among which, MIF-CUL1 axis was significantly increased. These results suggest that the limitations of vitamin D in SHPT treatment is closely related to oxyphil cell and may be attributed to the dysregulation of vitamin D transport and ubiquitin regulation of oxyphil cell.

Significance: Secondary hyperparathyroidism in end stage renal patients is one of the major challenges nephrology field faces. Emerging data indicate that oxyphil cell may participate in the pathophysiology of secondary hyperparathyroidism, while both calcimimetics and vitamin D receptor activators treatments are underperformed in controlling oxyphil cell proliferation. In the present study, we validated that the proliferation of oxyphil cells is associated with calcitriol treatment, and discovered that oxyphil cell nodules were significantly different from chief cells nodules in protein expression of mitochondria, protein synthesis and cell cycle regulation. It is noteworthy that DBP was downregulated in oxyphil nodules on protein level and may therefore participate in the resistance of calcitriol therapy by reducing the vitamin D transport capacity. Wnt signaling, TGF-β, ubiquitin mediated proteolysis and cell cycle pathways were significantly enriched in oxyphil cell nodules, among which, MIF-CUL1 axis may play an important role in the regulation of oxyphil proliferation and calcitriol resistance through ubiquitin mediated proteolysis. These results suggest that calcitriol treatment has limitations in oxyphil cell predominant SHPT, which may be attributed to the dysregulation of vitamin D transport and ubiquitin regulation of oxyphil cell, and the influence of microenvironment in uremia status may be the underlying reason.
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http://dx.doi.org/10.1016/j.jprot.2018.02.029DOI Listing
May 2018

Differentiation of minute virus of mice and mouse parvovirus by high resolution melting curve analysis.

J Virol Methods 2017 12 23;250:41-46. Epub 2017 Sep 23.

Guangdong Key Laboratory of Laboratory Animals, Guangzhou 510633, China. Electronic address:

Murine parvovirus is one of the most prevalent infectious pathogens in mouse colonies. A specific primer pair targeting the VP2 gene of minute virus of mice (MVM) and mouse parvovirus (MPV) was utilized for high resolution melting (HRM) analysis. The resulting melting curves could distinguish these two virus strains and there was no detectable amplification of the other mouse pathogens which included rat parvovirus (KRV), ectromelia virus (ECT), mouse adenovirus (MAD), mouse cytomegalovirus (MCMV), polyoma virus (Poly), Helicobactor hepaticus (H. hepaticus) and Salmonella typhimurium (S. typhimurium). The detection limit of the standard was 10 copies/μL. This study showed that the PCR-HRM assay could be an alternative useful method with high specificity and sensitivity for differentiating murine parvovirus strains MVM and MPV.
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http://dx.doi.org/10.1016/j.jviromet.2017.09.022DOI Listing
December 2017

Low employment and low willingness of being reemployed in Chinese working-age maintained hemodialysis patients.

Ren Fail 2017 Nov;39(1):607-612

f Department of Nephrology , Huashan Hospital Fudan University , Shanghai , China.

Aim: Returning to society plays an important role in improving the quality of life in maintenance hemodialysis (MHD) patients, and retention of employment is one of the core enablers. The study is to assess the employment status and to determine the variables for unemployment in Chinese MHD patients.

Methods: Prevalent MHD patients from four dialysis centers in Shanghai China were enrolled. We assessed patients' employment status, current social functioning, hemodialysis modality, annual income and general health condition. Among current unemployed working-age patients, the reasons of quitting jobs and willingness of being reemployed were evaluated.

Results: A total of 231 patients were studied, among which 114 patients were unemployed 1 year before hemodialysis. Among 117 employed patients, 16 patients quitted jobs before dialysis inception, while 49 patients quitted jobs at the initiation of HD, and 26 patients followed after a few months' HD. The main reasons for ceasing employment were physical insufficiency, conflict between dialysis and work schedules, lack of support from employers and resistance from family members. Among the 166 patients who were in their working age, 26 patients were employed. The unemployed patients had the characters of elder age, lower education level, higher annual family income, higher female ratio, lower blood flow, lower physical functioning, and social functioning and lower frequency of weekend hemodialysis and HDF/HF. Among the 140 unemployed patients, only 47 patients had the willingness of being reemployed. Their unemployment status was positively associated with elder age ((OR) 3.13, 95% CI, 1.08-9.1), lower education level ((OR) 1.97, 95% CI, 1.05-5.92), and higher family income ((OR) 7.75, 95% CI, 2.49-24.14).

Conclusion: Ratio of employment and willingness of being reemployed was low in MHD working-age patients. Lack of social and family's support also hampered patient's returning to society except for the HD treatment quality.
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http://dx.doi.org/10.1080/0886022X.2017.1361834DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6446148PMC
November 2017

Management of non-adherence to fluid intake restrictions in hemodialysis patients in a tertiary hospital: a best practice implementation project.

JBI Database System Rev Implement Rep 2016 08;14(8):309-22

1School of Nursing, Fudan University, Shanghai, China 2The Fudan Evidence Based Nursing Center: a Joanna Briggs Institute Centre of Excellence, Fudan University, Shanghai, China 3Hemodialysis Center, Huashan Hospital, Shanghai, China 4The Joanna Briggs Institute, Faculty of Health Sciences, The University of Adelaide, Adelaide, Australia.

Background: Non-adherence to fluid-intake restrictions is one of the most common problems for hemodialysis (HD) patients. A combined approach that involves patients, healthcare professionals and caregiver inputs based on best practice is important for enhancing adherence within a busy health system.

Objectives: The aim of this project was to promote evidence-based practice in the management of fluid-intake restrictions among HD patients in a HD center.

Methods: Six evidence-based criteria developed by the Joanna Briggs Institute were used as a basis for audits undertaken in the Hemodialysis Center of Huashan Hospital, Shanghai, mainly focusing on nurse education, fluid-intake management by patients and the role of caregivers in assisting with fluid-intake management by patients. The project included three phases and was conducted over 5 months. The Joanna Briggs Institute Practical Application of Clinical Evidence System and Getting Research into Practice audit tools for promoting change in health practice were used to examine compliance with the criteria before and after the implementation of best practice.

Results: Results from pre- and post implementation audits indicated that the compliance rates of four criteria reached 100% after the implementation of various strategies. One criterion (patients' self-monitoring) resulted in a compliance rate of 73.33%. One other criterion (patients' medical documentation) was maintained at 100% compliance. Patients' adherence to fluid intake, knowledge and attitude to self-management also improved significantly in the post implementation audit.

Conclusion: This project achieved a significant improvement in evidence-based practice for the management of non-adherence to fluid-intake restrictions in HD patients. An increase in the number of HD patients found to be adherent to fluid-intake restrictions was reported during this process.
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http://dx.doi.org/10.11124/JBISRIR-2016-003046DOI Listing
August 2016

Development of a duplex real-time RT-PCR for the simultaneous detection and differentiation of Theiler's murine encephalomyelitis virus and rat theilovirus.

J Virol Methods 2016 10 7;236:139-146. Epub 2016 Jul 7.

Guangdong Key Laboratory of Laboratory Animals, Guangdong Laboratory Animals Monitoring Institute, Guangzhou, China. Electronic address:

Theiler's murine encephalomyelitis virus (TMEV) and rat theilovirus (RTV), the member of the genus Cardiovirus, are widespread in laboratory mice and rats, and are potential contaminants of biological materials. Cardioviruses infection may cause serious complications in biomedical research. To improve the efficiency of routine screening for Cardioviruses infection, a duplex real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay was developed for simultaneous detection and differentiation of TMEV and RTV. The duplex assay was specific for reference strains of TMEV and RTV, and no cross-reaction was found with seven other rodent viruses. The limits of detection of both TMEV and RTV were 4×10(1) copies RNA/reaction. Reproducibility was estimated using standard dilutions, with coefficients of variation <3.1%. 439 clinical samples were evaluated by both duplex real-time RT-PCR and conventional RT-PCR. For 439 clinical samples,95 samples were positive for TMEV and 72 samples were positive for RTV using duplex real-time RT-PCR approach, whereas only 77 samples were positive for TMEV and 66 samples were positive for RTV when conventional RT-PCR was applied. Mixed infections were found in 20 samples when analyzed by conventional RT-PCR whereas 30 samples were found to be mixed infection when duplex real-time RT-PCR was applied. This duplex assay provides a useful tool for routine health monitoring and screening of contaminated biological materials of these two viruses.
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http://dx.doi.org/10.1016/j.jviromet.2016.07.004DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7119687PMC
October 2016

Low-dose versus high-dose heparin locks for hemodialysis catheters: a systematic review and meta-analysis.

Clin Nephrol 2016 Jul;86(7):1-8

Background: Low-dose heparin lock has been suggested as an effective and safe catheter lock in hemodialysis. However, whether a low-dose lock is superior to a high-dose heparin lock in preventing catheter-related infections and maintaining catheter patency is inconclusive.

Study Design: A systematic review and meta-analysis was performed by searching in NGC (National Guideline Clearinghouse), Cochrane Library, Joanna Briggs Institute Library, MEDLINE, EMbase, RNAO (Registered Nurses' Association of Ontario), Nursing Consult, PubMed, OVID, China Biology Medicine disc (CBMdisc), China National Knowledge Infrastructure (CNKI), Chongqing VIP (CQVIP), Wanfang Data Knowledge Service Platform, and major nephrology journals.

Setting And Population: Patients receiving hemodialysis with central venous catheters.

Selection Criteria For Studies: Randomized controlled trials (RCT), controlled clinical trials (CCT), and cohort studies comparing low-dose heparin lock with high-dose heparin lock in hemodialysis patients with central venous catheters. Intervention (exposure): Locking central venous catheters with low-dose heparin lock.

Outcomes: Primary outcomes include bleeding-related complications and catheter-related infections. Secondary outcomes include catheter retention time, catheter thrombosis/occlusion incidence, and catheter dysfunction.

Results: Five RCTs and three CCTs were eventually identified for the meta-analysis, with 916 patients in total. Meta-analysis showed that, compared with the high-dose heparin group (≥ 5,000 U/mL), the low-dose (< 5,000 U/mL) heparin lock could significantly reduce the incidences of bleeding- related complications RR = 3.29, 95% CI (2.19, 4.94), p < 0.00001) and catheterrelated infections (RR = 1.66, 95% CI (1.01, 2.73), p = 0.04). However, no significant differences were observed in the catheter retention time MD = 9.32, 95% CI (-6.70, 25.35), p = 0.25), catheter thrombosis/occlusion incidence (RR = 0.68, 95% CI (0.28, 1.65), p = 0.39), or catheter dysfunction (RR = 1.07, 95% CI (0.75, 1.53), p = 0.71) between the high- and low-dose heparin groups.

Conclusion: Low-dose heparin lock could decrease the incidences of catheter-related infections and bleeding-related complications without influencing the catheter retention time or the incidence of catheter thrombosis/occlusion or catheter dysfunction. Low-dose heparin lock solution should be recommended for the hemodialysis patients using central venous catheter in clinic.
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http://dx.doi.org/10.5414/CN108701DOI Listing
July 2016

Anemia management trends in patients on peritoneal dialysis in the past 10 years.

Int J Clin Exp Med 2015 15;8(10):18050-7. Epub 2015 Oct 15.

Department of Nursing, Huashan Hospital, Fudan University Shanghai 200040, P. R. China.

Objective: This study is to investigate anemia management trends in patients on peritoneal dialysis in Huashan Hospital in the past 10 years.

Methods: A total of 463 cases of patients receiving peritoneal dialysis from July 2004 to March 2014 were analyzed retrospectively. The patients enrolled in this study all were elder than 18 years old, those who had been followed up for at least 1 quarter. Clinical data of anemia including hemoglobin, serum iron, iron saturation, dosage of erythropoietin (EPO) and iron supplements were collected quarterly and then analyzed those data.

Results: In the first period of 25 quarters, in a certain range, increased dose of erythropoietin and iron content, can make the patient's hemoglobin gradually increased to the ideal level, which helps to treat the symptoms of anemia patients. Results of comparation before and after the education program showed that with the enhance the EPO injection compliance and treat anemia with the proper dose of EPO, the PD patients have stable level of Hemoglobin, accompanying with the decreased dose of oral Iron and intravenous iron.

Conclusion: It was shown that hemoglobin value was constantly stable. Treat anemia with proper dose of EPO individually could cure anemia effectively with the reducing of iron supplement improvement peritoneal dialysis patients' anemia related knowledge, and the good compliance of injection of EPO, serum iron and iron saturation could still keep stable, although declined intake of iron supplements. Therefore, optimizing anemia management in peritoneal dialysis patients is helpful to improve their quality of life.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4694300PMC
January 2016

Phosphate removal model: an observational study of low-flux dialyzers in conventional hemodialysis therapy.

Hemodial Int 2012 Jul 23;16(3):363-76. Epub 2012 Feb 23.

Division of Nephrology, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai, China.

Precise assessing phosphate removal by hemodialysis (HD) is important to improve phosphate control in patients on maintenance HD. We reported a simple noninvasive model to estimate phosphate removal within a 4-hour HD. One hundred sixty-five patients who underwent HD 4 hours per session using low-flux dialyzers made of polysulfone (1.2 m(2)) or triacetate (1.3 m(2)) were enrolled. Blood flows varied from 180 to 300 mL/min. Effluent dialysate samples were collected during the 4-hour HD treatment to measure the total phosphate removal. Predialysis levels of serum phosphate, potassium, hematocrit, intact parathyroid hormone, total carbon dioxide (TCO(2)), alkaline phosphatase, clinical and dialysis characteristics were obtained. One hundred thirty-five observations were randomly selected for model building and the remaining 30 for model validation. Total amount of phosphate removal within the 4-hour HD was mostly 15-30 mmol. A primary model (model 1) predicting total phosphate removal was Tpo(4)  = 79.6 × C(45) (mmol/L) - 0.023 × age (years) + 0.065 × weight (kg) - 0.12 × TCO(2) (mmol/L) + 0.05 × clearance (mL/min) - 3.44, where C(45) was phosphate concentration in spent dialysate measured at the 45 minute of HD and clearance was phosphate clearance of dialyzer in vitro conditions offered by manufacturer's data sheet. Since the parameter TCO(2) needed serum sample for measurement, we further derived a noninvasive model (model 2):Tpo(4)  = 80.3 × C(45)  - 0.024 × age + 0.07 × weight + 0.06 × clearance - 8.14. Coefficient of determination, root mean square error, and residual plots showed the appropriateness of two models. Model validation further suggested good and similar predictive ability of them. This study derived a noninvasive model to predict phosphate removal. It applies to patients treated by 4-hour HD under similar conditions.
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http://dx.doi.org/10.1111/j.1542-4758.2012.00678.xDOI Listing
July 2012

Effect of short-term low-protein diet supplemented with keto acids on hyperphosphatemia in maintenance hemodialysis patients.

Blood Purif 2011 7;31(1-3):33-40. Epub 2010 Dec 7.

Division of Nephrology, Huashan Hospital, Fudan University, Shanghai, China.

Aim: To evaluate the effects of short-term restriction of dietary protein intake (DPI) supplemented with keto acids on hyperphosphatemia in maintenance hemodialysis (MHD) patients.

Methods: Forty MHD patients with uncontrolled hyperphosphatemia were randomized to either low DPI with keto acid-supplemented (sLP) or normal DPI (NP) group for 8 weeks. After 8 weeks, the sLP group was shifted to NP for another 8 weeks. Low-protein diet (LPD) was individualized with total caloric intake 30-35 kcal/kg/day, protein intake of 0.8 g/kg/day and phosphate intake of 500 mg/day. Keto acids were supplied in a dosage of 12 pills per day. Calcium phosphorous metabolism index and nutritional index (serum albumin, total protein, somatometric measurements, 3-day diaries and Mini-Nutritional Assessment score) were recorded. C-reactive protein, CO(2) combining power and Kt/V were measured to evaluate the inflammation, metabolic acidosis and dialysis adequacy, respectively.

Results: Serum phosphorus level and calcium-phosphate product were significantly decreased at the end of the first 8 weeks in the sLP group compared to the basal value and the NP group (p < 0.001). No difference was observed in C-reactive protein, Kt/V and nutritional index, while CO(2) combining power was significantly higher at week 8 in the sLP group (p < 0.001).

Conclusion: Short-term restriction of DPI supplemented with keto acids could decrease hyperphosphatemia and calcium-phosphate product, while keeping stable nutritional status among MHD patients.
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http://dx.doi.org/10.1159/000321376DOI Listing
May 2011