1.Validation of the detection method for residual human coagulation factor Ⅺ in human prothrombin complex
Yong LIU ; Yurong YU ; Long YANG ; Zexiu LI ; Yao ZHANG ; Jing DENG ; Dan LI ; Yunhua CHEN ; Xuemei ZHAO
Chinese Journal of Blood Transfusion 2023;36(9):818-822
【Objective】 To establish an enzyme-linked immunosorbent assay (ELISA) method for the determination of residual human coagulation factor Ⅺ in human prothrombin complex and validate the method. 【Methods】 Human factor Ⅺ was reacted with the capture antibody coated on the microtiter plate. After appropriate washing steps, biotinylated primary antibody was bound to the captured protein. Excess primary antibody was washed away and bound antibody was reacted with horseradish peroxidase conjugated streptavidin. TMB substrate was used for color development at 450 nm. The dilution reliability, accuracy, specificity, repeatability, intermediate precision, linearity, range and durability were verified. 【Results】 The verification results showed that the accuracy and specificity of this method met the experimental requirements, with an average recovery rate of 109.2% and RSD of 6.93%. The repeatability RSD was 6.78%, and the intermediate precision RSD was 6.75%, indicating good precision. The linear regression correlation coefficient of standard curve was 0.999 9, showing good accuracy and precision within the linear range. The durability was verified by the incubation time and the validity period of reagent kit opening. The results showed that the RSD of the incubation time change was 6.62%, indicating that the incubation time of this detection method was controlled between 28 to 32 minutes, and there was no significant impact on the results. The RSD of the detection results before and after the reagent kit was opened and stored under conditions for 7 days was 3.84%, indicating that the preservation of the reagent kit according to the conditions for 7 days after opening has no effect on the FⅪ detection results. Both indicated that the method had good durability. The dilution reliability results showed that there was a "hook" effect in the detection of FⅪ residue in human prothrombin complex, which could be solved by diluting 100 to 200 times. 【Conclusion】 This method can be used for the determination of FⅪ residues of human prothrombin complex in laboratory.
2.Related substances in purified alliin determined by HPLC-MS/MS.
Yao-Zuo YUAN ; Jie GU ; Tai-Jun HANG ; Jian CHEN ; Zheng-Xing ZHANG
Acta Pharmaceutica Sinica 2007;42(6):639-642
To study the related substances in purified alliin, HPLC-MS/MS method carried out on a Phenomenex NH2 column was used for screen and identification of the related substances with full scan MS spectra determination of their [M + H] + ions and then the analyses of the retention time, product MS spectra and/or chemical reference standards. The full scan HPLC-MS chromatogram showed that there were seven major related substances in the purified alliin and their m/z of the [M + H] + ions with increasing retention were 116, 133, 147, 152, 175, 178 and 178, separately. And they were identified as proline, asparagine, glutamine, methiin, arginine, isoalliin and cycloalliin (both were isomers of alliin), respectively. The major related substances in purified alliin are amino acids, homologen and the isomers of alliin.
Chromatography, High Pressure Liquid
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methods
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Cysteine
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analogs & derivatives
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analysis
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Tandem Mass Spectrometry
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methods
3.Study on maternal periodontal diseases of the relationships between porphyromonas gingivalis, serum pro-immflamatory mediators and preterm low birth weight.
Yao LIN ; Zong-rui TIAN ; Hong-bo CHEN ; Bao-jun TAI ; Han JIANG ; Min-quan DU
West China Journal of Stomatology 2009;27(6):595-598
OBJECTIVETo investigate the associations between periodontal diseases, presence of Porphyromonas gingivalis (P. gingivalis), serum levels of pro-inflammatory mediators and preterm low birth weight (PLBW).
METHODS60 women (30 PLBW and 30 healthy women), were recruited after postpartum within 3 days in this case-control study. Periodontal measurements including plaque index (PI), bleeding index (BI), probing pocket depth (PD) and clinical attachment loss (CAL) were assessed. The subgingival plaque was collected before periodontal examination and analysed by polymerase chain reaction (PCR) for the presence of the 16S rRNA gene specific to P. gingivalis, while the venous and umbilical cord blood specimens collected were determined by enzyme-linked immunosorbent assay (ELISA).
RESULTSThe PLBW cases had a poorer oral conditions and the presence of P. gingivalis was found in a higher proportion in the PLBW than the healthy pregnant (56.7% vs. 30.0%, P < 0.05). Both of the presence of periodontitis and P. gingivalis have been found to be associated weakly with a shorter gestational age and a lower birth weight (P < 0.05). The levels of IL-1beta, IL-6 and prostaglandin-E2 (PGE2) in both of the blood samples showed higer levels between the PLBW and normal groups (P < 0.01). The IL-1beta and PGE2 levels in maternal serum were higher with a severe periodontal disease in the PLBW group (P < 0.01).
CONCLUSIONThere may be a possible link between periodontal diseases and PLBW.
Adult ; Case-Control Studies ; Dental Plaque ; Dental Plaque Index ; Female ; Humans ; Infant, Low Birth Weight ; Infant, Newborn ; Interleukin-6 ; Periodontal Diseases ; Periodontitis ; Porphyromonas gingivalis ; Pregnancy ; Premature Birth ; RNA, Ribosomal, 16S
4.Noninfectious fever following aortic surgery: incidence, risk factors, and outcomes.
Yun-tai YAO ; Li-huan LI ; Qian LEI ; Lei CHEN ; Wei-peng WANG ; Wei-ping CHEN
Chinese Medical Sciences Journal 2009;24(4):213-219
OBJECTIVETo determine the incidence, course, potential risk factors, and outcomes of noninfectious fever developed in patients after aortic surgery.
METHODSpatients who received operation for aortic aneurysm or dissection in our center from January 2006 to January 2008 were reviewed. Patients who met one of the following criteria were excluded: having a known source of infection during hospitalization; having a preoperative oral temperature greater than or equal to 38.0 degrees C; undertaking emergency surgery; having incomplete data. Univariate analysis was performed in patients with noninfectious postoperative fever and those without, with respect to demographics, intraoperative data, etc. Risk factors for postoperative fever were considered for the multivariate logistic regression model if they had a P value less than 0.10 in the univariate analysis.
RESULTSTotally 463 patients undergoing aortic surgery were enrolled for full review. Among them, 345 (74.5%) patients had noninfectious postoperative fever, the other 118 (25.5%) patients didn't develop postoperative fever. Univariate analysis demonstrated that several risk factors were associated with the development of noninfectious postoperative fever, including weight, surgical procedure, minimum intraoperative bladder temperature, temperature upon intensive care unit (ICU) admission, discharge, and during ICU stay, as well as blood transfusion. In a further multivariate analysis, surgical site of thoracic and thoracoabdominal aorta (odds ratio: 4.861; 95% confidence interval: 3.029-5.801; P=0.004), lower minimum intraoperative bladder temperature (odds ratio: 1.117; 95% confidence interval: 1.01-1.24; P=0.04), and higher temperature on admission to the ICU (odds ratio: 2.57; 95% confidence interval: 1.28-5.18; P=0.008) were found to be significant predictors for noninfectious postoperative fever. No difference was found between the febrile and afebrile patients with regard to postoperative hospitalization duration (P=0.558) or total medical costs (P=0.896).
CONCLUSIONNoninfectious postoperative fever following aortic surgery is very common and closely related with perioperative interventions.
Adult ; Aged ; Aneurysm, Dissecting ; surgery ; Aortic Aneurysm ; surgery ; Female ; Fever ; diagnosis ; epidemiology ; etiology ; Humans ; Incidence ; Interleukin-6 ; blood ; Male ; Middle Aged ; Postoperative Complications ; epidemiology ; etiology ; Risk Factors ; Transfusion Reaction
5.Evaluate of the therapeutic efficiency of the Chinese traditional medicine Gansu granule on chronic hepatitis B.
Xiao-rong CHEN ; Hua YAO ; Yin JIANG ; Shan-ming WU ; Xiao-fang ZHU ; Xia-qiu ZHOU ; Yi-mei CAI ; Yun-hui ZHUO ; Jian-jie CHEN ; Ling-tai WANG
Chinese Journal of Hepatology 2004;12(1):50-50
Adolescent
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Adult
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Aged
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Female
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Hepatitis B, Chronic
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drug therapy
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Humans
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Male
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Medicine, Chinese Traditional
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Middle Aged
6.Generation of a new strain of NOD/SCID/IL2Rγ mice with targeted disruption of Prkdc and IL2Rγ genes using CRISPR/Cas9 system.
Ya-Chen LIU ; Qu CHEN ; Xing-Long YANG ; Qing-Shuang TANG ; Kai-Tai YAO ; Yang XU
Journal of Southern Medical University 2018;38(6):639-646
OBJECTIVEThe NOD/SCID/IL2Rγ (NSG) mouse strain is the most widely used immunodeficient strain for xenograft transplantation. However, the existing SCID mutation is a spontaneous mutation of the Prkdc gene, which leads to leaky T cell developmental block and difficulty in genotyping. It is therefore important to develop a new strain of NSG mice with targeted disruption of Prkdc and IL2Rγ genes.
METHODSTargeted disruption of Prkdc and IL2Rγ genes was achieved using the CRISPR/Cas9 system. By intercrossing the knockout and NOD mice, we obtained a novel strain of NOD/SCID/IL2Rγ(NSG) mice, denoted as cNSG (Chinese NSG) mice.
RESULTSIn addition to the NOD mutation, cNSG mice exhibited a complete absence of T cells, B cells and NK cells. cNSG mice allowed more efficient engraftment of human cancer cells than the commonly used immunodeficient nude mice.
CONCLUSIONcNSG mice will provide an important xenotransplantation model for biomedical research.
7.Effect of patient decision aids on choice between sugammadex and neostigmine in surgeries under general anesthesia: a multicenter randomized controlled trial
Li-Kai WANG ; Yao-Tsung LIN ; Jui-Tai CHEN ; Winnie LAN ; Kuo-Chuan HUNG ; Jen-Yin CHEN ; Kuei-Jung LIU ; Yu-Chun YEN ; Yun-Yun CHOU ; Yih-Giun CHERNG ; Ka-Wai TAM
Korean Journal of Anesthesiology 2023;76(4):280-289
Background:
Shared decision making using patient decision aids (PtDAs) was established over a decade ago, but few studies have evaluated its efficacy in Asian countries. We therefore evaluated the application of PtDAs in a decision conflict between two muscle relaxant reversal agents, neostigmine and sugammadex, and sequentially analyzed the regional differences and operating room turnover rates.
Methods:
This multicenter, outcome-assessor-blind, randomized controlled trial included 3,132 surgical patients from two medical centers admitted between March 2020 and August 2020. The patients were randomly divided into the classical and PtDA groups for pre-anesthesia consultations. Their clinicodemographic characteristics were analyzed to identify variables influencing the choice of reversal agent. On the day of the pre-anesthesia consultation, the patients completed the four SURE scale (sure of myself, understand information, risk-benefit ratio, encouragement) screening items. The operating turnover rates were also evaluated using anesthesia records.
Results:
Compared with the classical group, the PtDA group felt more confident about receiving sufficient medical information (P < 0.001), felt better informed about the advantages and disadvantages of the medications (P < 0.001), exhibited a superior understanding of the benefits and risks of their options (P < 0.001), and felt surer about their choice (P < 0.001). Moreover, the PtDA group had a significantly greater tendency to choose sugammadex over neostigmine (P < 0.001).
Conclusions
PtDA interventions in pre-anesthesia consultations provided surgical patients with clear knowledge and better support. PtDAs should be made available in other medical fields to enhance shared clinical decision-making.
8.Dystrophin expression in mdx mice after bone marrow stem cells transplantation.
Shan-wei FENG ; Cheng ZHANG ; Xiao-li YAO ; Mei-juan YU ; Jing-lun LI ; Song-lin CHEN ; Tai-yun LIU ; Xi-lin LU
Acta Academiae Medicinae Sinicae 2006;28(2):178-181
OBJECTIVETo investigate the dynamic changes of dystrophin expression in mdx mice after bone marrow stem cells transplantation.
METHODSThe bone marrow stem cells of C57 BL/6 mice (aged 6 to 8 weeks) were injected intravenously into the mdx mice (aged 7 to 9 weeks), which were preconditioned with 7Gy gamma ray. The amount of dystrophin;expression in gastrocnemius was detected by immunofluorescence, reverse transcription-polymerase chain reaction and Western blot at week 5, 8, 12 and 16 after transplantation.
RESULTSAt week 5 after bone marrow stem cells transplantation, the dystrophin expression detected in mdx mice were very low; however, its expression increased along with time. At week 16 week, about 12% muscle cells of all transplanted mice expressed dystrophin. There were less centrally placed myonuclei than the control mdx mice, whereas peripheral myonuclei increased.
CONCLUSIONSAfter having been injected into mdx mice, the allogenic bone marrow stem cells have a trend to reach the injured muscle tissues and differentiate to fibers that can express dystrophin and the expression increased with time. The bone marrow stem cells participates in the repair and regeneration of the injured tissues permanently and constantly.
Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; Disease Models, Animal ; Dystrophin ; biosynthesis ; Hematopoietic Stem Cell Transplantation ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Inbred mdx ; Muscular Dystrophy, Duchenne ; metabolism ; surgery ; Transplantation, Homologous
9.An area CCD-based detection system for up-converting phosphor immune lateral flow assay
Chao LI ; Zi-Jiang YANG ; Yao-Hua DU ; Tai-Hu WU ; Feng CHEN
Military Medical Sciences 2017;41(11):906-911
Objective To make portable the detector for up-converting phosphor (UCP) immune lateral flow assay portable and increase the test precision by developing an area CCD-based detection system for UCP immune lateral flow assay.Methods The excitation light source was a 980 nm and 300 mW laser diode.In order to decrease the error induced by the un-uniformity of laser irradiation,a uniformity mirror was inserted in the beam and a calibration algorithm was optimized.The residual error from the un-uniformity irradiation was less than 1%.Results Thanks to the adjustability of the exposure time,the dynamic range of detection of the system was as high as 106 dB.The repeat test error for the very low signal was 1% (variation coefficient).The linear correlation coefficient between the tested T/C value and sample concentration was 0.998.Conclusion Compared to is traditional instrument,this detection system is static,portable and highly precise.
10.Research progress of single-particle focusing device and related technologies
yang Zheng DONG ; Feng CHEN ; hua Yao DU ; Zhi CHENG ; Jing WEI ; hu Tai WU
Chinese Medical Equipment Journal 2017;38(11):109-112,117
The present situation and research progress of the single-particle focusing device and related technologies were introduced in foreign countries and China. The principles and advantages of different single-particle focusing devices were analyzed from the aspects of liquid and gas single-particle focusing, and it's pointed out that rapid detection of the particles such as bacteria could be realized by liquid and gas single-particle focusing measures. The two measures both had brilliant prospects providing the structure and design were optimized to ensure high detection precision and efficiency, which could be promoted for bio-agents detection and fulminating infectious diseases prevention and treatment.