1.Determination of six catechins in Xinnaojian Capsules (Tablets) by QAMS
Jing XIE ; Jie WEI ; Luwei ZHOU ; Jinlian HE ; Leilei DU
Chinese Traditional Patent Medicine 2017;39(3):523-527
AIM To establish a quantitative analysis of multi-components by single marker(QAMS) method for the content determination of six catechins in Xinnaojian Capsules (Tablets) (tea extract).METHODS The analysis of 50% methanol extract of this drug was performed on a 35 ℃ thermostatic Shimadzu Wonda Cract ODS-2 column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of 0.5% acetic acid (A)-acetonitrile (B) flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 280 nm.With epigallocatechin gallate as an internal standard,the relative correction factors of epigallocatechin,catechin,epicatechin,gallocatechin gallate and epicatechin gallate were calculated,from which the content determination was made.RESULTS Six constituents showed good linear relationships within their own ranges (r ≥ 0.999 8),whose average recoveries were 96.00%-98.47% with the RSDs of 2.09%-2.91%.The results obtained by QAMS method approximated those obtained by external standard method.CONCLUSION This simple and reliable method can be used for the quality control of Xinnaojian Capsules (Tablets).
2.Protective Effect of Tibetan Medicine Compound Duoxuekang Capsules on Acute Lung Injury in Mice In-duced by Lipopolysaccharide
Xiaoxin HU ; Qiang LUO ; Leilei DU ; Yi ZHANG ; Silang JIANGYONG
China Pharmacy 2017;28(7):926-929
OBJECTIVE:To investigate the protective effect of Tibetan medicine Compound Duoxuekang capsules on the acute lung injury in mice induced by lipopolysaccharide. METHODS:60 KM mice were randomly divided into blank group,model group,dexamethasone group (positive control,1 mg/kg) and Compound Duoxuekang high-dose,medium-dose,low-dose groups (3.6,1.8,0.9 g/kg,calculated by crude drug),10 in each group. Intragastrically administrated,mice in blank group and model group intragastrically administrated equal volume of normal saline,once a day. After 7 d of administration,except for blank group, mice in other groups intraperitoneally injected lipopolysaccharide to induce acute lung injury. After 6 h modeling,pathological changes in the lung tissue was observed,lung tissue water content was measured,superoxide dismutase(SOD),glutathione peroxi-dase(GSH-Px),malondialdehyde(MDA)levels in lung tissue and IL-6,TNF-α levels in serum were detected. RESULTS:Com-pared with blank group,mice in model group showed pathological changes in congestion and edema,inflammatory cell infiltra-tion,obvious widened alveolar septum and alveolar wall;water content in lung tissue and IL-6,TNF-αlevels in serum were signifi-cantly increased (P<0.01);SOD,GSH-Px levels in lung tissue were significantly decreased (P<0.01). Compared with model group,injury degree of lung tissue reduced to varying degrees in each treatment group,except for the water content,MDA level in lung tissue and TNF-α level in serum in Compound Duoxuekang capsules low-dose group;the above-mentioned indexes in other groups were significantly improved (P<0.05 or P<0.01). CONCLUSIONS:Compound Duoxuekang capsules can obviously re-duce mice's oxidative stress and inflammatory response,and has certain protective effect on acute lung injury in mice.
3.Effects of Candida albicans on the expression of tumor necrosis factor-α and activation of the intracellular signaling molecule p38MAPK in a human acute monocytic leukemia cell line THP-1
Zhimin DUAN ; Leilei DU ; Rong ZENG ; Yongnian SHEN ; Suquan HU ; Weida LIU ; Qing CHEN ; Min LI
Chinese Journal of Dermatology 2015;(8):535-538
Objective To investigate the effects of Candida albicans on the expression of tumor necrosis factor-α(TNF-α)and activation of the intracellular signaling molecule p38 mitogen-activated protein kinase(p38MAPK)in a human acute monocytic leukemia cell line THP-1. Methods Some THP-1 cells were divided into several groups in vitro: two C. albicans groups treated with 105 CFU/ml and 106 CFU/ml heat-killed C. albicans respectively, a lipopolysaccharide (LPS)group treated with 100 μg/L LPS, a blank control group treated with RPMI 1640 medium, two dexamethasone-inhibited groups pretreated with 40 μg/L dexamethasone for 30 minutes followed by treatment with 106 CFU/ml heat-killed C. albicans and LPS respectively. After treatment for 1, 3 and 6 hours, real-time fluorescence-based quantitative PCR was performed to measure TNF-α mRNA expression in THP-1 cells in the above groups. Enzyme-linked immunosorbent assay(ELISA)was conducted to determine the level of TNF-α protein in the supernatant of THP-1 cells treated with 106 CFU/ml heat-killed C. albicans, 100 μg/L LPS or RPMI 1640 medium(blank control group)for 24 hours. Western blot was performed to measure the protein expression of p38MAPK and phosphorylated p38MAPK in THP-1 cells after treatment with 106 CFU/ml heat-killed C. albicans or RPMI 1640 medium (blank control group)for 30 and 60 minutes. Statistical analysis was carried out by using two-way analysis of variance, one-way analysis of variance and the least significant difference(LSD)-t test. Results Significant differences were observed in the mRNA expression level of TNF-α among the C. albicans groups, LPS group and blank control group (F = 110.98, P < 0.001). The mRNA expression level of TNF-α in THP-1 cells increased over time in a time-dependent manner after C. albicans treatment, with significant differences among different time points (F = 701.680, P < 0.001). Compared with the blank control group, both 106-CFU/ml C. albicans group and LPS group showed a significant increase in TNF-α protein expression (6385.70 ± 533.99 ng/L and 3212.06 ± 353.00 ng/L vs. 147.10 ± 0.53 ng/L, P < 0.001 and 0.005, respectively). An obvious increase was observed in the expression level of phosphorylated p38MAPK protein, but no significant changes were noted in that of p38MAPK protein, in THP-1 cells treated with 106 CFU/ml C. albicans for 30 and 60 minutes compared with the blank control group. The mRNA expression level of TNF-α significantly decreased in dexamethasone-pretreated 106-CFU/ml C. albicans group and LPS group compared with those without dexamethasone pretreatment(3.77 ± 0.62 vs. 208.50 ± 10.50, 6.20 ± 1.93 vs. 161.35 ± 1.65, both P < 0.001). Conclusions Heat-killed C. albicans can induce the activation of p38MAPK in and secretion of TNF-α by human THP-1 cells, which then participate in the innate immune response against C. albicans.
4.Study on the antiulcer effects of Veronicastrum axillare on gastric ulcer in rats induced by ethanol based on tumor necrosis factor-α (TNF-α) and endothelin-1 (ET-1)
Yong DU ; Weichun ZHAO ; Leilei LU ; Jiayan ZHENG ; Xishi HU ; Zhehan YU ; Lixin ZHU
Asian Pacific Journal of Tropical Biomedicine 2013;(12):925-930
Objective:To assess whether Veronicastrum axillare (V. axillare) can ameliorate ethanol-induced gastric mucosal lesions in rats, reduce the production of pro-inflammatory cytokines, suppress apoptosis and improve local microcirculation disturbances.
Methods:Totally 48 male Sprague-Dawley rats were randomly divided into six groups, eight rats in each group. Rats in the normal group and the model group were administered with 0.9%normal saline respectively. Rats in the positive group and ranitidine group were administered with 0.18% ranitidine suspension by intragastric administration respectively. Those in the high dose V. axillare group, the medium dose V. axillare group and the low dose V. axillare group were administrated with V. axillare at the daily dose of 2.8 g/kg, 1.4 g/kg and 0.7 g/kg by intragastric administration. Gastric mucosal lesions were produced by intragastric administration of absolute ethanol. Water extract of V. axillare was successively injected for 14 d and last day was injected 1 h before ethanol administration. Gastric mucosal ulcer index and ulcer inhibitory rate were counted by improved Guth methods. The tissue sections were made for pathological histology analysis. Also, we measured the concentrations of tumor necrosis factor-α (TNF-α) and endothelin-1 (ET-1) in gastric mucosal, as an index of the pro-inflammatory cytokines, apoptosis and local microcirculation. Besides, the mRNA contents of TNF-αand ET-1 were measured to verify effects on gene expression by real-time fluorescent quantitative PCR.
Results: Water extract of V. axillare significantly ameliorated the gastric mucosal lesions induced by ethanol administration (P<0.01). Pro-inflammatory cytokines, TNF-αand ET-1 were increased after ethanol administration and significantly reduced by water extract of V. axillare. The expressions of TNF-αand ET-1 mRNA were also be inhibited by water extract of V. axillare.
Conclusion: Current evidences show water extract of V. axillare is effective for defending against ethanol-induced gastric mucosal lesions, significantly inhibiting the production of pro-inflammatory cytokines and the expressions of TNF-αand ET-1 mRNA, which may be useful for inhibiting apoptosis and improving local microcirculation.
5.Contents Determination of Catechins Active Components in Lipid-lowering Slimming Health Products by Quantitative Analysis of Multi-Components via Single Marker
Jing XIE ; Jie WEI ; Luwei ZHOU ; Jinlian HE ; Qianlong WANG ; Leilei DU
China Pharmacy 2017;28(18):2529-2532
OBJECTIVE:To establish the method for contents determination of catechins active components in lipid-lowering slimming health products. METHODS:HPLC method was adopted. Using epigallocatechin gallate(EGCG)as a reference,relative correction factor(RCF)of EGCG to gallocatechin(EGC),catechin(C),epicatechin(EC),gallocatechin gallate(GCG)and gal-loylepicatechin(ECG)were calculated. The contents of EGC,C,EC,GCG and ECG in 5 batches of samples were calculated through RCF. The contents of EGC,C,EC,GCG and ECG determined by external standard method were used as measured value. The similarity of the value determined by external standard method with the value calculated by quantitative analysis of multi-com-ponents via single marker method(QAMS)was evaluated with vector included angle cosine method. RESULTS:The linear ranges of EGCG,EGC,C,EC,GCG and ECG were 0.0065-0.1305 mg/mL(r=0.9998)、0.0005-0.0107 mg/mL(r=0.9997)、0.0020-0.0400 mg/mL(r=0.9999)、0.0153-0.3053 mg/mL(r=0.9998)、0.0008-0.0155 mg/mL(r=0.9998)、0.0040-0.0792 mg/mL (r=0.9999);RSDs of precision,stability and reproducibility tests were all lower than 2.0%.The recoveries were 95.07%-100.35%(RSD=1.94%,n=6)、95.24%-101.87%(RSD=2.79%,n=6)、96.08%-103.86%(RSD=3.01%,n=6)、97.51%-101.06%(RSD=1.45%,n=6)、96.01%-101.66%(RSD=2.27%,n=6)、96.20%-102.89%(RSD=2.71%,n=6),respectively. There was no signifi-cant difference between measured value and calculated value. CONCLUSIONS:The method is simple,precise,stable and repro-ducible,and can be used for contents determination of catechins active components in lipid-lowering slimming health products.
6.Chromosomal abnormalities with male infertility
Yuan DONG ; Yuting JIANG ; Richeng DU ; Jing WU ; Leilei LI ; Ruizhi LIU
Chinese Journal of Urology 2013;(6):410-413
Objective To describe different types of chromosomal abnormalities on male infertility.Methods From May 2006 to May 2012,2034 infertile males with genetic counseling underwent chromosome karyotype analysis,semen routine examination and reproductive hormones levels detection.The data from them were analyzed.Results 267 cases of chromosomal abnormalities were detected in 2034 cases (13.13%).258 cases underwent semen routine examination in 267 cases with chromosomal abnormalities,of which 190 cases of azoospermia,58 cases of oligozoospermia,10 cases of semen normal.In 267 cases of chromosomal abnormalities,including 169 cases (63.30%) of number abnormalities,mainly with azoospermia,157 cases of Klinefelter syndrome (KS) (58.80%),7 cases of 47,XYY (2.62%),4 cases of Turner syndrome (1.50%),1 case of marker chromosome (0.37%) ; 49 cases (18.35%) of structural abnormalities mainly with oligozoospermia,including 32 cases of chromosomal translocations (11.99%),17 cases of inversion (6.37%) ; 4 cases of sex reversal (1.50%) with azoospermia; 45 cases of chromosome polymorphism (16.85%) mainly with oligozoospermia.Non-mosaicism KS patients' age,testicular volume,semen volume,and serum reproductive hormones levels were compared between different groups of semen results,and there were no significant difference except age.Conclusions Chromosome abnormalities were the most important genetic causes of abnormal semen quality and male infertility.It is necessary to be performed chromosome karyotype analysis for infertile males.
7.Analysis of Y chromosome abnormalities and azoospermia factor microdeletions in male infertility
Yuan DONG ; Jing WU ; Richeng DU ; Yuting JIANG ; Leilei LI ; Ruizhi LIU
Chinese Journal of Laboratory Medicine 2013;(1):50-52
Objective To study the relationship between Y chromosome abnormalities and AZF microdeletions in males with reproductive failure.Methods A case-control study was conducted in 2694 reproductive failure men with age ranges from 23 to 49 years old from the Institute of Reproductive Medicine of Jilin Province.Patients were divided into three groups:spermatogenic failure group (n =1332),disadvantage pregnancy outcomes group (n =994) and adverse birth outcomes group.All patients underwent chromosomal karyotype analysis (G-banding).AZF microdeletions were further investigated in patients with Y chromosomal abnormalities by PCR.The Chi-square test was used to compare the frequency of Y chromosome abnormalities in three groups.Results Of the 51 cases of Y chromosome abnormalities (1.89%,51/2694),32 were (2.40%,32/1332) in the spermatogenic failure group,15 were (1.51%,15/994) in disadvantage pregnancy outcomes group and 4 were (1.09%,4/368) in adverse birth outcomes group.There was no significant difference in Y chromosome abnormalities among different groups (x2 =3.895,P >0.05).AZF microdeletions were detected in 10 cases (19.61%,10/51) of Y chromosome abnormalities patients with spermatogenic failure.Conclusions The incidence of Y chromosomal abnormalities in three reproductive failure groups is similar.Chromosome karyotype analysis and AZF microdeletions examination could identify the genetics etiology in males with reproductive failure.
8.The application of operating room quality backward system in instrument place management.
Hui DU ; Anjie HE ; Leilei ZENG
Chinese Journal of Medical Instrumentation 2010;34(5):384-385
OBJECTIVEImprovement of the surgery instrument's clean quality, the optimized preparation way, reasonable arrangement in groups, raising the working efficiency.
METHODWe use the quality backward system into the instrument clean, the pack and the preparation way's question, carry on the analysis and the optimization, and appraise the effect after trying out 6 months.
RESULTSAfter finally the way optimized, instrument clean quality distinct enhancement; The flaws in the instrument clean, the pack way and the total operating time reduce; the contradictory between nurses and the cleans arising from the unclear connection reduces, the satisfaction degree of nurse and doctor to the instrument enhances.
CONCLUSIONSUsing of operating room quality backward system in the management of the instrument clean, the pack and the preparation way optimized, may reduce flaws in the work and the waste of human resources, raise the working efficiency.
Materials Management, Hospital ; organization & administration ; Operating Rooms ; organization & administration ; standards ; Quality Control ; Software
9.Effects of the yeast form of Sporothrix schenckii on activation of p38MAPK and expression of interleukin-6 in human THP-1 macrophage-like cells
Caixia LIU ; Leilei DU ; Zhimin DUAN ; Rong ZENG ; Yongnian SHEN ; Suquan HU ; Weida LIU ; Qing CHEN ; Min LI
Chinese Journal of Dermatology 2017;50(8):562-566
Objective To evaluate effects of the yeast form of Sporothrix schenckii on activation of p38 mitogen-activated protein kinase (p38MAPK) and expression of interleukin-6 (IL-6) in macrophagelike THP-1 cells,which were differentiated from the human acute monocytic leukemia cell line THP-1.Methods THP-1 macrophage-like cells were divided into 3 groups to be treated with the yeast form of Sporothrix schenckii at a concentration of 2 × 106 colony-forming units (CFU)/ml (yeast form group),100 mg/L curdlan (curdlan group) and RPMI 1640 medium (blank control group) respectively.Real-time fluorescence-based quantitative PCR was performed to measure the mRNA expression of IL-6 in THP-1 macrophage-like cells in the above 3 groups after 3-and 6-hour treatment separately,and enzyme-linked immunosorbent assay (ELISA) to detect the level of IL-6 in the culture supernatant of THP-1 macrophagelike cells after 24-hour treatment.Western blot analysis was conducted to determine the protein expression of p38MAPK and phosphorylated p38MAPK (p-p38MAPK) in the above 3 groups after 30-and 60-minute treatment separately.Other THP-1 macrophage-like cells were pretreated with 100 nmol/L dexamcthasonc (a p38MAPK inhibitor) for 30 minutes,and then were divided into 3 groups to be treated with the yeast form of Sporothrix schenckii,curdlan and RPMI 1640 medium respectively,and changes in the level of pp38MAPK and mRNA expression of IL-6 were also detected.Statistical analysis was carried out with SPSS19.0 software by using one-way or multi-way analysis of variance and least significant difference (LSD) test.Results Significant differences in the mRNA expression of IL-6 in THP-1 macrophage-like cells were observed among the yeast form group,curdlan group and blank control group (F =5 552.22,P <0.001) after 3-hour treatment (56.81 ± 7.36,26.69 ± 1.22 and 0.97 ± 0.05,respectively) and 6-hour treatment (378.03 ± 16.67,276.24 ± 39.13 and 1.02 ± 0.04,respectively).Additionally,the yeast form group showed significantly higher mRNA expression of IL-6 after 6-hour treatment than that after 3-hour treatment (q =16.74,P < 0.001).After 24-hour treatment,the level of IL-6 in the culture supernatant of THP-1 macrophage-like cells also significantly differed among the yeast form group,curdlan group and blank control group (59.96 ± 18.16 pg/L,91.01 ± 17.27 pg/L,5.50 ± 2.30 pg/L,respectively;F =26.62,P < 0.01),and was significantly higher in the yeast form group than in the blank control group (P < 0.01).After 30-and 60-minute treatment,the protein expression of p-p38MAPK was significantly higher in the yeast form group than in the blank control group (both P < 0.01).Moreover,the mRNA expression of IL-6 (4.46 ± 1.03 vs.493.52 ± 113.87,P < 0.001) and protein expression of p-p38MAPK (2.29 ± 0.37 vs.4.55 ±0.46,q =10.81,P < 0.01) were both significantly lower in the yeast form group with dexamethasone pretreatment than in that without dexamethasone pretreatment.Conclusion In vitro treatment with the yeast form of Sporothrix schenckii can enhance the expression of IL-6 in human THP-1 macrophage-like cells by activating the p38MAPK signaling pathway.
10.Preliminary clinical analysis of concurrent chemoradiotherapy combined with nituzumab for inoperable locally advanced esophageal cancer
Cheng ZENG ; Dan YANG ; Rongxu DU ; Leilei JIANG ; Xin DONG ; Dongming LI ; Rong YU ; Huiming YU ; Anhui SHI
Chinese Journal of Radiation Oncology 2021;30(4):353-356
Objective:The standard treatment for inoperable locally advanced esophageal cancer is concurrent chemoradiotherapy, but the survival was not satisfied. Nituzumab is a humanized IgG monoclonal antibody against EGFR. The purpose of this study is to investigate the toxicity and efficacy of concurrent chemoradiotherapy combined with nituzumab for locally advanced esophageal cancer.Methods:We retrospectively reviewed the clinical data of locally advanced esophageal cancer who were treated with concurrent chemoradiotherapy combined with nituzumab in Peking University Cancer Hospital from June 2015 to June 2020. Kaplan- Meier method was used for analysis. Results:Thirty Patients were enrolled this study.After a median follow-up of 22.5 months, The objective response rate was 93%. The 1-year, 2-year, 3-year overall survival rates were 83%, 57% and 41%, with the progression-free survival rates 75%, 47% and 32%, with the local-recurrence free survival rates 83%, 53% and 37%, with the metastasis-free survival rates 75%, 51% and 36%, respectively.The incidence of grade≥3 hematological toxicity was 32%. There were 16% patients experiencing grade≥3 esophagitis.Conclusion:The preliminary result of concurrent chemoradiotherapy combined with nituzumab is effective and safe for patients with locally advanced esophageal cancer.