1.THE COMPARISON OF THREE KINDS OF ENZYMELINKED IMMUNOSORBENT ASSAY ON SCREENING THE “LEAKY” PHENOTYPE SCID MICE
Yi LI ; Heng CUI ; Xue YE ; Yu YAO
Acta Anatomica Sinica 1953;0(01):-
Objective Choose an easy method to screen the “leaky" phenotype SCID mice before experimental use. Methods IgG in SCID mice serum was assayed by three kinds of ELISA(sandwich ELISA, sandwich indirect ELISA and Avidin\|biotin sandwich ELISA). Results Avidin\|biotin sandwich ELISA is the most sensitive method of the three, and 41 SCID mice of 6~8 weeks old were assayed with this method. The average IgG of 40 SCID mice was 0 24?0 16mg/L, only one was 2\^3mg/L. The “Leaky" probability was 2\^4%.Conclusion\ The Avidin\|Biotin sandwich ELISA is an easy, sensitive and reliable method to screen“leaky" SCID mice.\;[
2.Re-evaluation of interval debulking surgery in advanced epithelial ovarian cancer
Yu MA ; Yi LI ; Heng CUI ; Xudong LIANG ; Zhijian TANG ; Xiaoping LI ; Yan ZHAO ; Lihui WEI
Chinese Journal of Obstetrics and Gynecology 2012;47(5):355-360
Objective Previous study showed that interval debulking surgery (IDS) may improve the survival of patients with advanced epithelial ovarian cancer (EOC).The precise significance of IDS needs to be evaluated.Methods Totally 136 consecutive patients with stage Ⅲ c or Ⅳ EOC (including primary peritoneal carcinoma and primary fallopian tube carcinoma ) who completed primary debulking surgery (PDS) and platinum-based chemotherapy were enrolled from January 2000 to December 2009 in a retrospective cohort study.The study group was divided into three groups:65 cases underwent optimal PDS (Group A ),41 cases received chemotherapy alone after suboptimal PDS (Group B ),and 30 patients underwent IDS after suboptimal PDS (Group C).All patients received six to eight courses of platinum-based combination chemotherapy (paclitaxel plus carboplatin/cisplatin,cyclophosphamide plus epirubicin and cisplatin).Patients' clinical characteristics,perioperative situation and prognosis were compared. Results Sixty-five cases (47.8%,65/136) from 136 patients achieved optimal PDS.For Group C,77% (23/30)patients obtained optimal debulking surgery after IDS.Intraoperative injury rates were similar between Group B and Group C ( P > 0.05 ).Mild perioperative complications rate was also similar ( P >0.05 ).Median progression-free survival (PFS) of Group A was 26 months.Median overall survival (OS) of Group B and Group C were 3l months and 40 months,respectively (P =0.254).Median PFS of Group B and Group C were 13 months and 24 months,respectively (P =0.289).Although when it came to 20 months after PDS,patients who underwent IDS had a significantly lower progressive disease (PD) rate (Group B 33% versus Group C 61%,P =0.046 ),it still showed that there was no significant difference in either OS or PFS of these two groups.Those patients in Group C who obtained no visible residual got similar PFS (27 months) comparing to Group A (26 months,P =0.730),but OS was still shorter (P =0.010).Conclusions For advanced EOC patients,IDS has little effect on improving survival.While it is safe and acceptable,also may prolong PFS in those patients who got no visible residual after IDS.The results suggest that IDS might be used as an alternative treatment for advanced EOC patients who cannot obtain optimal PDS in certain local hospitals.
3.Effect of Autoantibody Tests in Diagnosis of Autoimmune Diseases in Children
qian, ZHANG ; xiao-chuan, WANG ; jing-yi, WANG ; ye-heng, YU
Journal of Applied Clinical Pediatrics 2006;0(21):-
Objective To study the effect of autoantibody test which includes antinuclear antibodies(ANA),antinuclear antibody fluorescence patterns,anti-extractable nuclear antigen(ENA) antibodies and anti-double strands DNA(ds-DNA) antibodies in the diagnosis of pediactic autoimmune diseases.Methods Of all the inpatients which had positive results of autoantibody tests,135 cases were reviewed.The autoantibody assay,positive value(PV) analysis were performed respectively.Results PV of ANA test to autoimmune diseases was 0.36 which was proportional to the intensity of fluorescence;Of all the fluorescence patterns,speckled(fine) had a relatively high PV;Anti-ENA and anti-dsDNA antibody tests had higher PV than ANA test.Conclusion Fluorescence intensity,(anti-ENA) antibody test and anti-dsDNA antibody test may be useful in identifying autoimmune diseases in clinic.
5.Clinical and laboratory features of B-cell chronic lymphocytic leukemia: a retrospective analysis of 503 cases
Heng LI ; Wenjie XIONG ; Huimin LIU ; Shuhua YI ; Rui LYU ; Tingyu WANG ; Zhen YU ; Lugui QIU ; Zengjun LI
Journal of Leukemia & Lymphoma 2017;26(6):325-330
Objective To investigate the clinical and laboratory characteristics of patients with chronic lymphocytic leukemia (CLL). Methods 503 patients with CLL admitted from October 1998 to February 2015 were retrospectively analyzed. Baseline characteristics were compared using Chi-square test and Kaplan-Meier methodology was undertaken for survival analyses. Results The median age was 58 years (26-86 years):335 cases were male and 168 cases were female. 204 cases (40.5%) were at the clinical stage of Binet A, followed by Binet B (148 cases, 30.1%) and Binet C (151 cases, 29.3%). 108 cases (21.1%) had anemia at diagnosis, while 113 cases (26.5 %) had an elevated level of lactate dehydrogenase and the expression of CD38 was detected among 100 cases (29.1 %). Clonal abnormalities were observed using fluorescence in situ hybridization (FISH) analysis. Those involving 13q deletion were the most frequent (156 cases, 47.3 %), followed by IgH translocation (22.4 %), trisomy 12 (21.2 %) and 17p deletion (14.5 %). The mutational status of immunoglobulin heavy chain variable region was determined among 230 cases, 165 cases (71.7%) of which were found to be with mutated status. The most frequently encountered gene was V4-34 (28 cases, 12.4 %). The median progression-free survival (PFS) was 89.0 months (95 %CI 75.0-103.0 months), while the median overall survival was 129.0 months (95 %CI 106.9-151.1 months). Conclusion Compared with patients in the western world, CLL patients in this study are younger at diagnosis and have longer overall survival, which, to some extent, could reflects the characteristics of CLL patients in China.
6.Bifidobacterium DNA upregulates Th1 type response of umbilical cord blood mononuclear cell.
Hui ZHAO ; Xiao-chuan WANG ; Jing-yi WANG ; Ye-heng YU ; Chuan-qing WANG ; Yi YANG
Chinese Journal of Pediatrics 2006;44(6):415-419
OBJECTIVETo study the effect of bifidobacterium genomic DNA on umbilical cord blood mononuclear cell (CBMC), and investigate the immunoregulation of bifidobacterium DNA and explore possible mechanisms by which bifidobacterium acts against allergic reaction.
METHODSBifidobacterium genomic DNA (bDNA) and human DNA (hDNA) were extracted with phenol/chloroform/isoamyl alcohol and stored at -20 degrees C for later use. Parts of bDNA were completely digested with DNaseI (d-bDNA) at 37 degrees C. CBMCs were isolated with Ficoll from umbilical cord blood and incubated at 37 degrees C in a 5% CO2 humidified incubator. These cells were divided into four groups, control group: without any stimulant; bDNA group: stimulated with 25 microg/ml bDNA; d-bDNA group: stimulated with 25 microg/ml d-bDNA; hDNA group: stimulated with 25 microg/ml hDNA. The cells were stimulated with different stimulants in vitro, at the end of incubation culture supernatant and cells were collected. IL-12 and IL-10 levels in the culture supernatant were measured by enzyme linked immuno sorbent assay (ELISA); cells secreting IL-4 and IFN-gamma were counted by enzyme linked immunospot (ELISPOT) assay; and total RNA was isolated from the cells to assay T-bet and GATA3 mRNA expression levels by reverse transcription polymerase chain reaction (RT-PCR).
RESULTSSix hours after stimulation there was no significant difference in IL-12 level in supernatant among the four groups; 12 hours after stimulation, IL-12 level in supernatant of bDNA treated group was significantly higher than that of each of the other groups, so were the results obtained at 24 hours and 48 hours after stimulation (P < 0.05). No significant difference could be detected in IL-12 level in supernatant among the other 3 groups. On the other hand, 6 hours after stimulation there was no significant difference in IL-10 level in supernatant among the four groups. But 12 and 24 hours after stimulation IL-10 level in supernatant of bDNA treated group was lower than that of each of the other groups, but the difference was not statistically significant. The count of IFN-gamma secreting cells of bDNA treated group was higher than that of the other groups, while IL-4 secteting cells of bDNA treated group were lower than that of the other groups. After bDNA stimulation, nuclear factor T-box expressed in T cells (T-bet) mRNA expression was conspicuously enhanced as compared to the other three groups (P < 0.05). GATA3 mRNA transcription in CBMC had no significant change after bDNA stimulation.
CONCLUSIONbDNA could promote secretion of Th1 type cytokine IL-12, while Th2 type cytokine IL-10 level of cell supernatant was decreased. bDNA could stimulate secretion of IFN-gamma by CBMC and inhibit secretion of IL-4. T-bet mRNA expression was highly enhanced after bDNA stimulation. bDNA could upregulate Th1 type response, which may be one of important mechanisms by which bifidobacterium inhibit allergic response.
Bifidobacterium ; cytology ; genetics ; Cell Culture Techniques ; DNA, Bacterial ; biosynthesis ; metabolism ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Fetal Blood ; cytology ; immunology ; GATA3 Transcription Factor ; genetics ; Humans ; Infant, Newborn ; Interferon-gamma ; immunology ; secretion ; Interleukin-10 ; immunology ; secretion ; Interleukin-12 ; immunology ; secretion ; Interleukin-4 ; immunology ; secretion ; Leukocytes, Mononuclear ; immunology ; secretion ; RNA, Messenger ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; T-Box Domain Proteins ; genetics ; Th1 Cells ; drug effects ; immunology ; secretion
7.Effect of environment temperature on the recovery of microdialysis probe for monoamine neurotransmitter.
Heng-Yi ZHANG ; Hong-Yi SHI ; En-Yu WANG
Journal of Zhejiang University. Medical sciences 2009;38(3):271-275
OBJECTIVETo investigate the effect of environment temperature on the recovery of microdialysis probe for neurotransmitters.
METHODSNeurotransmitters NE, DA, and 5-HT were dialyzed with 4 mm membrane length microdialysis probes in different environmental temperature. There were three conditions: lower temperature condition, the temperature of standard solution and that of the perfusate were both 24 degree; higher temperature condition,both were 37.5 degree; and the middle temperature:the perfusate was 24 degree and the standard solution was 37.5 degree. The concentrations of neurotransmitters in the dialyzed solution and the standard solution were analyzed by HPLC-ECD, and recoveries were then calculated.
RESULTIn lower temperature condition,the recoveries of microdialysis probe for NE, DA, 5-HT were 18.3 %, 19.6% and 16.9%, respectively. In middle temperature condition, the recoveries were 29.6%, 30.7% and 24.3%, respectively, and in higher temperature condition, those were 49.2%, 47.5% and 37.2%, respectively. With the analysis of variance, the recoveries for NE, DA, 5-HT increased with temperature significantly (P<0.01).
CONCLUSIONBoth the perfusate and the standard solution affects the environmental temperature of microdialysis probe, which in turns affects the recovery of microdialysis probe for neurotransmitters. So in order to calculate the recovery more accurately, the standard solution/the perfusate should be kept in body temperature.
Animals ; Biogenic Monoamines ; analysis ; Brain ; metabolism ; Brain Chemistry ; Chromatography, High Pressure Liquid ; Humans ; Microdialysis ; methods ; Neurotransmitter Agents ; analysis ; Oxadiazoles ; Temperature
8.Surgical treatment of large or giant cavernous sinus hemangiomas via epidural approach.
Xin-guang YU ; Yi-heng YIN ; Ding-biao ZHOU
Chinese Journal of Surgery 2012;50(8):724-727
OBJECTIVETo summarize the curative effect of surgically treated large or giant cavernous sinus hemangiomas (CSH) via epidural approach.
METHODSFrom June 1999 to June 2011, 19 cases of CSH including 15 female and 4 male patients, ranging from 26 to 70 years (mean 45.3 years) were retrospectively reviewed. Ptosis/ocular motility disorders (10 cases), headache/ophthalmalgia (7 cases), decreased visual acuity (7 cases) and facial hyperesthesia (4 cases) were the most common presenting complaints. The epidural approach was taken and the tumor dissection was performed at the interval between trochlear nerve and ophthalmic nerve and the interval between ophthalmic nerve and maxillary nerve. The curative effect was followed up regularly.
RESULTSGross total resection was attained in 13 cases, near-total resection in 4 cases, subtotal resection in 1 case and partial resection in 1 case. The follow-up period was between 6-144 months (mean 41.5 months). The results of last follow-up were as follows: among the 10 patients with ptosis/ocular motility disorders, complete remission was achieved in 5, improvement in 2, no change in 2 and lost follow-up in 1. Facial hypoesthesia symptom got complete remission in 2 patients and partial remission in the other 2 patients. The other symptoms mentioned above were all relieved. One patient got complication with impairment of vision and ocular motility disorders.
CONCLUSIONSCombined with skilled skull base techniques, surgical treatment of large or giant CSH via epidural approach is an effective method and the preservation of cranial nerves could be attained to an acceptable level.
Adult ; Aged ; Cavernous Sinus ; pathology ; Female ; Follow-Up Studies ; Hemangioma ; surgery ; Humans ; Male ; Middle Aged ; Neurosurgical Procedures ; methods ; Retrospective Studies ; Treatment Outcome
10.Proteomic Analysis of Rat Brain Stem with DAI by MALDI-TOF-MS.
Guan-heng REN ; Ning-guo LIU ; Yi-jiu CHEN ; Yan SHI ; Dong-hua ZOU ; Ping HUANG ; Zheng-dong LI ; Ho Yu SHA ; Kai-fei DENG
Journal of Forensic Medicine 2016;32(1):13-17
OBJECTIVE:
To establish a diagnostic model for diffuse axonal injury (DAI) by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). To screen the proteins or peptides associated with DAI for providing the biomarkers with theoretic foundation.
METHODS:
Fifteen male Sprague-Dawley rats were randomly divided into DAI group (n = 10) and control group (n = 5). The protein or peptide expression profiles of rat brain stem were detected by MALDI-TOF-MS. ClinProTools 2.2 software was used to find specific peaks, and a diagnostic model was established by the genetic algorithm.
RESULTS:
There were significant differences in 61 peaks of DAI group (P < 0.05), 9 peaks were down-regulated and 52 up-regulated. The diagnostic model was established based on 5 different peaks. The specificity and sensitivity of cross validation was 96.14% and 95.98%; while the specificity and sensitivity of blind validation showed was 73.33% and 70.00%, respectively.
CONCLUSION
A specific and sensitive diagnostic model of DAI can be established by MALDI-TOF-MS to provide a potential value for determining DAI in forensic practice.
Animals
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Biomarkers
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Brain Stem/metabolism*
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Diffuse Axonal Injury/diagnosis*
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Down-Regulation
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Male
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Peptides/blood*
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Proteomics
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Rats
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Rats, Sprague-Dawley
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Sensitivity and Specificity
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Software
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods*
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Up-Regulation