1.Development of LC-MS/MS assay for simultaneous determination of 25 psychotropic drugs in human blood
Chen XU ; Lei TANG ; Huidi JIANG
Chinese Journal of Forensic Medicine 2017;32(1):16-20
Objective To establish a LC-MS/MS assay for simultaneous determination of 25 psychotropic drugs in human blood and apply the method to analyze the relationship between(driving under the inference of drugs, DUID)and traffic violations. Methods Acetonitrile was added to the sample for precipitating the protein, the mixture was centrifuged for 10min at 4℃ , 14000r/min, the supernatant was filtered, and then the filtrate was analyzed with LC-MS/MS. The separation was performed on C18 chromatography column (50mm×3.0mm, 2.6μm), the mobile phase was consisted of 0.1% formic acid (phase A), acetonitrile:methanol=1:1 (phase B) with gradient elution. Results The calibration curves for 25 psychotropic drugs were linear over the ranges of 0.05~20ng/mL, R=0.9944~0.9996, the assay recoveries were 83.0%~99.7%, the method recoveries were 80.2%~97.4%, the intraday and interday precious were 1.6%~14% and 3.1%~14% respectively. The method was applied to analyze 3140 human samples stored by Center of Forensic Sciences of Hangzhou. Conclusion The method is sensitive and accurate for fast detection of 25 psychotropic drugs in human blood.
2.The role of ADME evaluation in translation research of innovative drug.
Yao LIU ; Lan HONG ; Lushan YU ; Huidi JIANG ; Jianzhong CHEN ; Qin MENG ; Shuqing CHEN ; Su ZENG
Acta Pharmaceutica Sinica 2011;46(1):19-29
New Chemical Entities (NCEs) development is a systematic long-term project that involves multiple disciplines. The translation research will help to build an advanced R&D system from the basic laboratory research, preclinical studies and clinical evaluation to clinical application of drug, for the purpose of shortening the R&D cycle and accelerate the launch of new drugs. In new drug R&D and its clinical application, drug disposition (absorption, distribution, metabolism, excretion, ADME) properties are important criteria for assessing drug-likeness of candidates. ADME evaluation of NCEs plays an important role in the translation research throughout innovative drug R&D process. Therefore, ADME evaluation at the early stage of drug design and development will be helpful to improve the success rate and reduce costs, and further access to safe, effective drugs.
3.Follow-up study of 1035 children with recurrent wheezing
Lin WEI ; Yue ZHANG ; Huidi JIN ; Beilan SAN ; Jie CHEN ; Jiong YU ; Mengbei XU
Journal of Clinical Pediatrics 2014;(6):532-535
Objective To investigate the long-term outcome of children with recurrent wheeze and to determine the effectiveness of inhaled hormone therapy. Methods One thousand and thirty-five children with recurrent wheezing were followed up for more than 4 years and the data were retrospectively evaluated. Results Of 1035 cases, 751 (72.56%) patients outgrew their wheeze during the follow-up period, whereas the other 284 (27.44%) patients had recurrence wheeze during the last two years. The age of wheezing onset was<3 years in 542 (52.37%) cases, from 3 to 7 years in 386 (37.29%) cases, and from 7 to 12 years in 107 (10.34%) cases. There was significant difference in clinical control rate among groups with different wheezing ages onset (χ2=45.27, P<0.001). Children with wheezing age onset from 7 to 12 years had the lowest clinical control rate. Among 1035 wheeze children, 343 (79.95%) children in 429 cases who received inhaled hormone therapy for more than one year outgrew their wheeze. Whereas 408 (67.35%) in 606 cases who did not receive inhaled hormone therapy outgrew their wheeze. There was significant difference of clinical control rate between inhaled group and non-inhaled group (P<0.01). Con-clusions The age of wheezing onset is<7 years in 89.66%of children with recurrent wheeze. Most of them can be clinicalycon-trolled. The long term inhaled hoemone therapy for children with recurrent wheeze can reduce the risk of developing adulthood asthma.
4.Human cytomegalovirus infection and its significance in peripheral blood leukocytes of patients with systemic lupus erythematosus
Jing CHEN ; Jiangbiao XIONG ; Wenjing YE ; Xiaochun ZHU ; Huidi ZHANG ; Lifang ZHANG ; Xiangyang XUE
Chinese Journal of Rheumatology 2014;18(3):199-202
Objective To detect the prevalence of human cytomegalovirus (HCMV) in peripheral blood of patients with systemic lupus erthematosus (SLE) and explore its role in the pathogenesis of SLE.Methods HCMV DNA was isolated from the peripheral blood leucocytes (PBLs) of 60 patients with SLE and 111 healthy controls.Nested polymerase chain reaction (nPCR) technology was used to investigate the gene of HCMV glycoprotein gB (UL55) in these specimens.HCMV infections in the PBLs of SLE patients were confirmed by HCMV-UL55 detection.Two-sample t test,nonparametric test,Chi-square test and Fisher probabilities were used to analyze.Results Agarose gel electrophoresis and sequencing analysis showed that established nPCR could specifically detect HCMV-UL55 gene,the HCMV infection rate was significantly higher in patients with SLE than in the healthy controls (P<0.01).Positive rates of HCMV infection in SLE group and controls were 41.7% (25/60) and 1.8% (2/111),respectively.Compared to the SLE patients with HCMV-negative PBLs,the positive rate of Rib-P [26%(9/35) vs 56%(14/25),x2=5.659,P=0.017],the positive rate of direct Coomb's test [37%(13/35) vs 72%(18/25),x2=7.096,P=0.008] and the level of antiβ2GP1 [21.3 (9.9,51.8) U/ml vs 13.6 (5.9,23.1) U/ml,U=2.017,P=0.044] were significantly higher than those in the SLE patients with HCMV-positive PBLs.Compared to the SLE patients with HCMV-negative PBLs,the number of red blood cells [(3.65±0.10)×1012/L vs (3.17±0.17)×1012/L,t=2.574,P=0.013] and lymphocytes [(1.37±0.14)×1012/L vs (0.90±0.13)×1012/L,t=2.456,P=0.017] in peripheral blood and the hemoglobin levels [(110±19) g/L vs (98±5)g/L,t=2.034,P=0.048] of the SLE patients with HCMV-positive decreased significantly.At the same time,the positive rate of hematuria [40%(14/35) vs 72%(18/25),x2=6.000,P=0.014] and 24 h proteinuria [0.80 (0.53,2.37)g vs 0.48 (0.13,1.21)g,U=2.140,P=0.032],which indicated kidney damage were also significantly increased in SLE patients with HCMV-positive PBLs.Conclusion The infection of HCMV in peripheral blood cells may take part in the development of SLE.
5. Analysis of Diagnosis and Treatment of Gastroesophageal Reflux Disease in A Hospital
Chinese Journal of Gastroenterology 2019;24(4):230-234
The incidence of gastroesophageal reflux disease (GERD) increased year by year and with a high risk of recurrence. The effect of proton pump inhibitors (PPIs) is not sufficient in some of the GERD patients. Aims: To investigate the status of diagnosis and treatment of GERD in a hospital. Methods: A total of 312 GERD patients from January 2017 to March 2018 at the 909th Hospital/the Affiliated Dongnan Hospital of Xiamen University were enrolled. Frequency of use of PPIs, initial therapy regimen, replacement of PPIs and dose of PPIs, use of antipsychotics, course of therapy and reexamination were retrospectively analyzed. Results: About 1/3 patients received the treatment of antipsychotics. The initial therapy was mainly PPIs+antacid or PPIs+antacid+prokinetics. PPIs was replaced in 51 patients, and the dose of PPIs was increased in 286 patients. And 54 patients received standard course of treatment. Conclusions: The treatment regimen of GERD is basically rational, however, health education should be strengthened and patient's compliance be elevated for increasing the treatment efficacy of GERD.
6.Construction of evaluation form with syndrome differentiation for lumbar disc herniation
Yuaner CHEN ; Xiaoyan XU ; Ying FENG ; Baoyu WANG ; Huidi CHENG
Chinese Journal of Nursing 2018;53(2):207-211
Objective To construct Evaluation Form for TCM Nursing with Syndrome Differentiation for Lumbar Disc Herniation in order to provide references for better nursing care of patients with lumbar disc herniation with syndrome differentiation.Methods According to the principle of "discern Yin and Yang,grasp the main symptoms",literature analysis and Delphi method were used to draft the primary and secondary symptoms of lumbar disc herniation's common syndromes,and Evaluation Form for TCM Nursing with Syndrome Differentiation for Lumbar Disc Herniation was developed.Totally 84 patients were recruited,and results evaluated by clinical physicians and nurses using the evaluation form were compared.Results By means of the evaluation form,80 patients were considered as the type of "Xue yu qi zhi",1 patient as the type of "Han shi bi zu",the others three as the type of "Gan shen kui xu";according to clinical physicians,83 patients were the type of "Xue yu qi zhi",and 1 patient was the type of "Gan shen kui xu".The consistency rate of two methods was 96.43%,and there was no significant difference (P>0.05).Conclusion There was no statistically significant difference between the results from clinical physicians and nurses using the evaluation form.The evaluation form is applicable in clinical practice,which can lay the foundation for nurses to carry out nursing with syndrome differentiation.
7.Constructing a recombinant adenovirus carrying human cytomegalovirus UL138 gene and evaluating the effects of UL138 gene on the functions of monocytes
Gangqiang GUO ; Jing CHEN ; Wenjing CHEN ; Lulu YE ; Xiangwei SUN ; Liang ZHANG ; Huidi ZHANG ; Qiao′ai LIN ; Xian SHEN ; Lifang ZHANG ; Xiangyang XUE
Chinese Journal of Microbiology and Immunology 2016;36(9):667-675
Objective To construct a recombinant adenovirus carrying UL138 gene, which was re-lated to the latent infection of human cytomegalovirus, and to investigate the effects of UL138 gene on the functions of THP-1 mononuclear cells. Methods The recombinant adenovirus expressing the UL138 gene was packaged. The titer of the recombinant adenovirus was determined by calculating 50% tissue culture in-fective dose ( TCID50 ) . THP-1 mononuclear cells were infected with the recombinant adenovirus at different multiplicity of infection (MOI) and the optimal MOI was determined (100 PFU/cell) by observing the ex-pression of green fluorescent protein ( GFP ) . Changes in the expression of proinflammatory cytokines by THP-1 mononuclear cells that was induced by overexpressed UL138 were analyzed by quantitative PCR. The expression of chemokines and their receptors were measured by quantitative PCR array. Results The re-combinant adenovirus carrying the UL138 gene was successfully constructed with a titer of 1×1011 PFU/ml. The rate of THP-1 mononuclear cells that was infected with the recombinant adenovirus was 60% at the MOI of 1 ∶ 100. Results of the RT-PCR analysis and Western blot assay further confirmed that the recombinant adenovirus could infect THP-1 mononuclear cells successfully and the expression of UL138 protein increased gradually over time. The overexpressed UL138 in THP-1 mononuclear cells significantly inhibited the expres-sion of IL-18, IL-1β, IL-6, IL-8 and TNF-α as indicated by the results of quantitative PCR. Results ob-tained from the quantitative PCR array analysis showed that most of the chemokines and their receptors were downregulated in the transfected THP-1 mononuclear cells except for the chemokines of CCL17, CCL21, CCL2 and XCL2 and the receptors of CCR2, CXCR1, CXCR2, CXCR4 and CX3CR1 which were upregulat-ed. Conclusion We successfully constructed the recombinant adenovirus carrying UL138 gene which could be used to infect THP-1 mononuclear cells. Overexpressed UL138 in THP-1 mononuclear cells significantly affected the functions of THP-1 mononuclear cells.
8.Long-term outcome and prognostic indicators of diffuse proliferative lupus nephritis
Tianxin CHEN ; Ying ZHOU ; Bo CHEN ; Huidi ZHANG ; Jianna ZHANG ; Xiaohan YOU ; Chaosheng CHEN
Chinese Journal of Rheumatology 2020;24(1):7-12
Objective:To investigate the long-term outcome and prognostic indicators of diffuse pro-liferative lupus nephritis (DPLN).Methods:The primary endpoint of long-term follow-up and factors pos- sibly influencing the outcome were analyzed retrospectively in DPLN patients admitted to the First Affiliated Hospital of Wenzhou Medical University between Jan 1, 2000 and Dec 31, 2014. Patients were classified into three groups according to the evaluated glomerular filtration rate(eGFR) on the first day of admission: eGFR≥60 ml·min -1·1.73 m -2 (regular illness group); 15 ml·min -1·1.73 m -2≤eGFR<60 ml·min -1·1.73 m -2 (serious illness group); eGFR<15 ml·min -1·1.73 m -2 or dialysis (critical illness group). Clinical, histological, and outcome differences among the three groups were evaluated and compared using one-way analysis of variance (ANOVA) , χ2 test, Kaplan-Meier survival curve and Cox reggression analysis. Results:167 DPLN patients were studied [155 women; mean age (30±10) years; mean follow-up of (61±45) months]. Renal and patient survival of all patients was 86% at 5 years and 79% at 10 years. Kaplan-Meier analysis showed the renal and patient survival rate at 10 years in the regular illness group, serious illness group and critical illness group was 91%, 70% and 8%, respectively ( χ2=121.93, P<0.01, overall); regular illness group vs serious illness group ( χ2=4.05, P<0.05); regular illness group vs critical illness group ( χ2=97.05, P<0.01); serious illness group vs critical illness group ( χ2=52.28, P<0.01). Multivariable Cox regression analysis found that haematoglobin (Hb)<80 g/L [ HR=2.7, 95% CI(1.2, 6.3), P=0.019], eGFR<60 ml·min -1·1.73 m -2 [ HR=4.1, 95% CI(2.0, 8.2), P<0.01] and large crescents ≥30%[ HR=1.8, 95% CI (1.1, 2.9), P=0.021], were risk factors for the long-term outcome. Conclusion:DPLN patients with normal or slightly decreased renal function have a better long-term prognosis. Moderate to severe impairment of renal function, anemia and large crescents are associated with poor outcome.