1.Biological characteristics of dendritic cells derived from peripheral blood of patients diagnosed with syphilis
Haiping ZHANG ; Wenming ZHAO ; Wei ZHU ; Shi LIAN
Chinese Journal of Dermatology 2008;41(5):304-306
Objective To study the biological characteristics of dendritic cells (DCs) derived from peripheral blood mononuclear cells (PBMCs) of patients diagnosed with syphilis. Methods PBMCs were isolated from 16 patients clinically and serologically diagnosed with syphilis, and from 16 healthy human controls, then cultured with GM-CSF and IL-4. On day 10, the monocyte-derived dendritic cells (MoDCs)of the patients and controls were collected and subjected to the detection of surface molecules by flow cytometry; TpN17 was used to stimulate MoDCs from the controls, the expression of phosphorylated ERK was detected by Westem blotting 20 minutes following the stimulation. Results The positivity rate of CD80 was significantly increased in the patients with syphilis than that in the controls (51.90% vs 33.67,P < 0.05), while no significant difference was observed in the expressions of CD83, CD86 or HLA-DR be tween the two groups (16.53% vs 15.99%, 66.13% vs 59.32%, 91.29% vs 90.51%, all P 0.05). The ex pressions of CD80 and CD83 on the surface of MoDCs were enhanced in a dose-dependent manner after ex-posure to TpN17. The expression of cytoplasmic phosphorylated ERK was observed in MoDC stimulated by TpN17, but not in those without the treatment. Conclusions Antigenic stimulation with Treponema pal-lidum may be a reason for phenotypic abnormality of MoDCs derived from patients with syphilis. TpN17 may stimulate the maturation of DCs through the ERK signal transduction pathway.
2.Performance evaluation and results analysis of chemiluminescence method for detecting plasma BNP
Xiaoming JIANG ; Wenming LIU ; Wen SHI ; Yinghui JIANG ; Jianhua XU
International Journal of Laboratory Medicine 2015;(24):3532-3534
Objective To analyze and evaluate the precision ,accuracy and linearity of the chemiluminescence method for detec‐ting plasma BNP .Methods According to the experimental schemes of CLSI EP15‐A2 ,EP6‐A files and other relevant documents , the precision ,accuracy and linearity of the Siemens Centaur XP chemiluminescence instrument for detecting BNP ,and the detection results were compared with the performance declared by manufacturer or the quality target formulated by laboratory .Results The imprecision of BNP detected by the chemiluminescence method was less than 1/3 TEa regulated by the Clinical Laboratory Center of Ministry of Health (allowable total error);the variation coefficient index (CVR) of internal quality control data was less than ± 2;the relative bias of the results of external quality control blind samples with the target values were less than Tea regulated by the Ministry of Health ;the linear evaluation results showed that BNP was once linearity in the range of 5 .3‐4696 .7 pg/mL .Conclusion The precision ,accuracy and linearity of the Siemens Centaur XP chemiluminescence instrument for detecting BNP can accord withthe quality objectives requirements and meet the clinical needs .
3.In vitro study and molecular modeling on 17-AAG-Ioaded polymethyl methacrylate bone cement for multiple myeloma
Xiangjun SHI ; Boran DU ; Xinru DU ; Wenming CHEN
Chinese Journal of Orthopaedics 2016;36(15):988-994
Objective To explore the possibility of polymethyl methacrylate (PMMA) bone cement as the carrier for lipophilic drugs through in vitro cytotoxicity study and molecular modeling with PMMA and 17-allylamino-17-demethoxy-geldanamycin (17-AAG) loaded PMMA bone cement.Methods The 17-AAG loaded bone cement was made by mixing method.In vitro antitumor activity with MTT assay for PMMA,17-AAG,the 24 h and 48 h released solution of 17-AAG loaded PMMA bone cement were evaluated.Through Material Studio 5.0,the interaction between 17-AAG and PMMA through the model of Amorphous Cell and energy optimization of Forcite was explored.Results The inhibition ratio of MMA for tumor cells is 9.21%±0.06% with 50 μmol/L.The 24 h released solution of 17-AAG loaded PMMA bone cement (17-AAG∶ PMMA=1∶ 4 000) inhibits the tumor cells 66.15%±0.43% which has a quick released influence on 17-AAG.The inhibition of 24 h released solution of 17-AAG-loaded PMMA bone cement (17-AAG∶PMMA=1 ∶ 1 000,1∶2 000) shows no significance compared with PMMA released solution (P<0.05).The 48 h released solution of 17-AAG-PMMA (17-AAG∶ PMMA=1∶ 1 000,1∶2 000,1∶4 000) inhibits U26630.25%±4.47%,30.24%±3.42%,50.52%±5.20%,with a significant difference with PMMA.The molecular model showed that the interaction between 17-AAG and PMMA was van der Waalz bonds,which drove 17-AAG inside or on the surface of PMMA bone cement.Conclusion PMMA bone cement can be used as a carrier for lipophilic drugs.It has antitumor activity and influences the release of 17-AAG with different ratio,for example it has a sustained-released influence on 17-AAG in 17-AAG-PMMA (17-AAG∶PMMA=1 ∶ 1 000,1∶2 000).Molecular model implies that 17-AAG exists inside or on the surface the PMMA bone cement through van der Waalz bonds.
4.Umbilical cord blood mononuclear cell transplantation for treatment of decompensated cirrhosis
Yuqin XU ; Wenming SHI ; Jinshun LI ; Jianguo XU ; Hongwen LI ; Xiang HU
Chinese Journal of Tissue Engineering Research 2013;(45):7975-7980
BACKGROUND:Orthotopic liver transplantation is the most effective therapy for the treatment of end-stage liver diseases, but the lack of donor source, immune rejection, and repeated infections limit its application. Stem celltransplantation technology provides a new idea for the treatment of end-stage liver diseases. A variety of methods have been confirmed to successful y induce umbilical cord blood mesenchymal stem cells converted into liver cells in vitro.
OBJECTIVE:To explore the clinical efficacy and feasibility of human umbilical cord blood mononuclear cells transplantation in the treatment of decompensated cirrhosis.
METHODS:Twenty-three patients with decompensated cirrhosis received al ogeneic human umbilical cord blood mononuclear celltransplantation. Serum alanine aminotransferase, albumin, cholinesterase, total bilirubin and prothrombin time were detected at post-transplantation weeks 2, 4, 8 and 24. Improvement in clinical signs and symptoms as wel as adverse reactions was observed.
RESULTS AND CONCLUSION:Liver function had no changes at 2 weeks after human umbilical cord blood mononuclear celltransplantation (P>0.05). At 4 weeks after celltransplantation, serum alanine aminotransferase was improved significantly (P<0.05), but the other indexes stil had no changes. Until 12 weeks after celltransplantation, there were significant improvements in al the liver function indicators (P<0.05) and the liver stiffness (P<0.05). By the end of 24 weeks, al the test results were improved significantly (P<0.01). Clinical symptoms were al eviated, including fatigue improvement in 20 cases (87%), improved appetite in 21 cases (91%), and relieved ascites in 19 cases (83%). No severe adverse reactions were found during the transplantation and 24-week fol ow-up. These findings suggest that human umbilical cord blood mononuclear cells transplantation is effective and safe for the treatment of decompensated cirrhosis, which can be considered as a clinical therapy for patients with advanced cirrhosis.
5.Dosimetric comparison and clinical application of RapidArc and intensity-modulated radiotherapy for postoperative radiotherapy of cervical cancer
Guozi YANG ; Zhenyu PAN ; Wenming XIA ; Yinghua SHI ; Huafang WANG ; Lihua DONG
Chinese Journal of Radiological Medicine and Protection 2014;34(1):37-40
Objective To compare the planning quality and acute toxicity between RapidArc and fixed gantry angle dynamic intensity modulated radiotherapy (IMRT) in the postoperative radiotherapy for cervical cancer patients.Methods All 35 patients with cervical cancer who had received postoperative radiotherapy were studied,including 17 patients with RapidArc and 18 patients with IMRT.All plans were prescribed 50 Gy in 25 fractions.The dose-volume histogram data,the conformity index and homogeneity index of the targets,the monitor units (MUs) and delivery time were compared.During the treatment,the incidence of acute intestinal and bladder side effects were also compared.Results Compared to IMRT,the conformity index of RapidArc was better(t =3.13,P < 0.05),but the homogeneity index was slightly worse (t =-4.25,P < 0.05).The V20 and V30 of femoral head planned by RapidArc was significantly lower than that by IMRT (t =2.56,2.34,P < 0.05).The mean MU for RapidArc was reduced by 52.1% compared with IMRT.The mean treatment time for RapidArc was decreased by 46.8% compared with IMRT.There was no difference in the incidence of acute intestinal and bladder toxicity between the two groups.Conclusion For patients with cervical cancer who need prophylactic postoperative radiotherapy,both RapidArc and IMRT plan can achieve equal target coverage and organs at risk(OAR) sparing.There is no significant difference in dosimetric parameters and acute toxicity between the two groups.Compared with IMRT,RapidArc plan has fewer MUs and less treatment time and significantly improves the treatment efficiency.
6.Phaeohyphomycosis caused by Cladosporium sphaerospermum: a case report
Qiuxia CHEN ; Wenming HUANG ; Wen LI ; Jianqiang SHI ; Fengyan BAI ; Shunfan LI
Chinese Journal of Dermatology 2010;43(12):823-825
A 53-year-old male patient was admitted to the hospital on March 26, 2006. Ten years prior to the presentation, a small furuncle developed on the dorsum of his right hand, and subsided after 2-week treatment with erythromycin ointment; one month later, a broadbean-sized cutaneous ulcer developed on the dorsum of the same hand. After anti-infective treatment, the ulcer healed while the lesional skin thickened, and long-term topical treatment with compound dexamethasone acetate cream showed no obvious effect Dermatological examination revealed an irregular verrucous plaque measuring 2.5 cm × 4 cm with little exudation on the dorsum of the right hand. KOH preparation of the skin lesion revealed brown spores. Sabouraud's dextrose agar culture grew restricted, velvety and dark green colony, and microscopy revealed branched, globular conidiophores generated by cladospores. DNA sequencing showed that the isolate was different from Cladosporium sphaerospermum (AB100654) by 2 bases in the sequence of D1/D2 region of 26S rDNA, from Cladosporium sphaerospermum (AY625063) by 5 bases in the sequence of internal transcribed spacer 1 and 2 (ITS 1 and ITS 2), but fully consistent with Cladosporium sphaerospermum (AM 176719) in the sequence of ITS region. The isolate was identified as Cladosporium sphaerospermum. Hematoxylin-eosin stain of the lesional tissue revealed granulomatous changes,and PAS stain demonstrated brown spores. A diagnosis of phaeohyphomycosis was made. Antifungal susceptibility testing indicated that the isolate was highly sensitive to itraconazole. The lesion obviously subsided after treatment with oral itraconazole 0.2 g once daily for 8 weeks, but the patient was lost to follow up 2 months later.
7.Construction and identification of interference plasmid targeting on TNFAIP8
Wenming LIU ; Jingjing YANG ; Ruyi HU ; Xingfeng QIU ; Chunyan SHI ; Zhongquan QI ; Zhongchen LIU ; Guohong ZHUANG
Chinese Journal of Immunology 2015;(5):650-654
Objective:To construct and screen the high efficiency interference plasmid of TFAIP8-shRNA-pSIREN-RetroQ.Methods:Selected and synthesized three Target Sequence of TNFAIP8 shRNA1,TNFAIP8 shRNA2,TNFAIP8 shRNA3,and construct the TNFAIP8 interference plasmid.Transfection TNFAIP8-shRNA-pSIREN-RetroQ interference plasmid to A549 cells.Filter out the highest interference efficiency plasmid by detecting the mRNA and protein levels using RT-PCR and Western blot methods.Results:We successfully design and built three TNFAIP8-shRNA-pSIREN-RetroQ interference plasmids,and screen out the highest efficiency interference plasmid.Conclusion: Three interference plasmids targeting the TNFAIP8 gene have been constructed successfully and provide a useful tool for studying the function of TNFAIP8.
8. Effect and related mechanism of miRNA-21 on hydrogen peroxide-induced C-kit+ cardiac stem cells apoptosis
Ranzun ZHAO ; Yan WANG ; Wenwen DENG ; Bei SHI ; Xianping LONG ; Zhenglong WANG ; Wenming CHEN
Chinese Journal of Cardiology 2017;45(11):971-977
Objective:
To explored the effect and related mechanism of miRNA-21 on hydrogen peroxide-induced C-kit+ cardiac stem cells apoptosis.
Methods:
C-kit+ cardiac stem cells were isolated from SD rats by the methods of enzyme digestion and magnetic bead. Cells were divided into the following experimental groups: (1) negative control mimics (NCM)group: cells were transfected with negative control miRNA-21 mimics for 48 hours; (2)mimics group: cells were transfected with miRNA-21 mimics for 48 hours; (3) NCM+ H2O2 group: negative control miRNA-21 mimics were transfected into cells for 48 hours and then treated with 100 μmol H2O2 for 2 hours; (4)mimics+ H2O2 group: miRNA-21 mimics were transfected into cells for 48 hours and then treated with 100 μmol H2O2 for 2 hours. mRNA of miRNA-21 was detected by RT-PCR. The apoptosis rate of C-kit+ cardiac stem cells was determined using the annexin V-FITC/PI staining assay. Western blot was employed to measure the expression of apoptosis related proteins(Caspase-3, Bax, and Bcl-2).
Results:
(1) Compared with the NCM group, the mRNA expression level of miRNA-21 was significantly up-regulated in mimics group, while obviously down-regulated in NCM+ H2O2 group(all
9.Clinical characteristics of acute myeloid leukemia patients with TET2 gene mutation and effects of TET2 mutation on therapeutic efficacy and prognosis
Wenxia SHI ; Yongxin GUO ; Junjie SHEN ; Wenming CHEN ; Wenwen GUO ; Yanyan TANG ; Shuige YANG ; Zhongliang SUN ; Daoping SUN
Journal of Leukemia & Lymphoma 2021;30(12):719-725
Objective:To investigate clinical features of adult patients with acute myeloid leukemia (AML) with TET2 gene mutation and effects of TET2 mutation on therapeutic efficacy and prognosis.Methods:A total of 123 newly diagnosed adult AML patients (except for acute promyelocytic leukemia) admitted to Jining No.1 People's Hospital from March 2017 to April 2021 were selected. Mutations of 24 AML-related genes including TET2 mutation were detected by using second-generation sequencing technology. Patients were divided into two groups according to the presence of TET2 mutation: TET2 mutation group and TET2 wild type group. The differences in clinicopathological characteristics, short-term efficacy and survival of both groups were compared.Results:Among 123 patients, TET2 mutation was detected in 28 cases (22.8%). Compared with TET2 wild type group, the patients were older [(59±15) years vs.(49±16) years, t = 2.984, P = 0.003], French-American-British (FAB) Corporative Group M 4 and M 5 subtypes were more common [75.0% (21/28) vs. 51.6% (49/95), χ2 = 4.838, P = 0.028], and the positive rate of CD34 in AML patients was lower in TET2 mutation group [46.4% (13/28) vs.72.6% (69/95), χ2 = 6.685, P = 0.010]. Moreover, TET2 mutation was more likely to be accompanied with ZRSR2 mutation [10.7% (3/28) vs. 1.1% (1/95), P = 0.037] and NPM1 mutation [35.7% (10/28) vs.17.9% (17/95), χ2 = 4.008, P = 0.045], but less likely to be accompanied with IDH1/2 mutation [0 vs.17.9% (17/95), P = 0.012]. However, there were no statistically significant differences in gender, peripheral blood leukocyte count at initial diagnosis, hemoglobin level, platelet count, bone marrow blasts ratio, cytogenetics and the European LeukemiaNet (ELN) risk stratification between the two groups (all P>0.05). In addition, there were no significant differences in the overall response rate (ORR) of 1 cycle chemotherapy [75.0% (12/16) vs. 66.7% (42/63), χ2 = 0.410, P = 0.522] and demethylation therapy [66.7% (4/6) vs. 44.4% (8/18), P = 0.640]. The difference in overall survival (OS) of both groups was not statistically significant [median OS time: 23 months (95% CI 5-41 months) vs. 35 months (95% CI 18-52 months, P = 0.498]. Conclusions:In AML patients, TET2 mutation is associated with advanced age, M 4 and M 5 subtypes, and low expression of CD34 on AML blasts. TET2 mutation is commonly accompanied by ZRSR2 and NPM1 mutation, but not IDH1 or IDH2 mutation. TET2 mutation may have no significant effects on therapeutic efficacy and survival in the whole cohort of AML patients without risk stratification.
10.Analysis of risk factors for multiple myeloma combined with radiation-based sarcopenia
Jie REN ; Lei QI ; Xingchen YAO ; Jinzhou WANG ; Xiangjun SHI ; Wenming CHEN ; Xinru DU
Chinese Journal of Orthopaedics 2023;43(9):567-573
Objective:To analyze the risk factors of radiation-based sarcopenia in patients with multiple myeloma (MM).Methods:A total of 185 clinical and imaging data of patients with MM admitted to Beijing Chaoyang Hospital from September 2009 to October 2019 were retrospectively analyzed. The area of the erector spinae muscle and the area of fatty infiltration (FI) in the fascial compartment were measured by Image-pro Ρlus software, and the area of the fat-free erector spinae muscle and the fat infiltration rate (FI%) were calculated. Sarcopenia was defined as an erector spinae area of less than 3 197 mm 2 in males and 2 895 mm 2 in females. The differences in gender, age, body mass index, disease duration, hemoglobin, leukocytes, platelets, albumin, serum calcium, lactate dehydrogenase, serum creatinine, alkaline phosphatase, M-protein, serum β 2-microglobulin, bortezomib chemotherapy, receipt of stem cell transplantation, osteopathy, stage, recurrence and progression of MM between the sarcopenia group and the normal muscle group were compared. Binary logistic regression was used to analyze the independent risk factors of sarcopenia in MM patients. Kaplan-Meier curves were drawn to compare the survival rates between the two groups. Results:53.0% (98/185) of MM patients were complicated with sarcopenia: there were 30 males, whose fat-free erector spinae area was 25.0±6.0 cm 2, the FI of erector spinae was 12.0±4.8 cm 2, and the FI% was 31.5%±12.0%, while there were 68 females, whose fat-free erector spinae area was 22.7±4.2 cm 2, the FI of erector spinae was 10.7±4.1 cm 2, and the FI% was 30.2%±9.8%. 47.0% (87/185) of MM patients had normal muscle mass: there were 62 males, whose fat-free erector spinae area was 40.6±6.5 cm 2, the FI of erector spinae was 9.3±4.8 cm 2, and the FI% was 17.9%±7.4%, while there were 25 females, whose fat-free erector spinae area was 33.6±5.1 cm 2, the FI of erector spinae was 9.9±3.0 cm 2, and the FI% was 21.9%±5.7%. There were statistically significant differences in the gender composition ratio (χ 2=30.47, P<0.001), hemoglobin ( t=-2.73, P=0.007), serum creatinine ( Z=-2.26, P=0.024), receipt of stem cell transplantation (χ 2=4.32, P=0.038), and MM recurrence and progression (χ 2=3.85, P=0.050) between the two groups. However, there were no significant differences in age, body mass index, course of disease, leukocytes, platelets, albumin, alkaline phosphatase, lactate dehydrogenase, serum calcium, M-protein, serum β 2-microglobulin, bortezomib chemotherapy, osteopathy or MM stage ( P>0.05). Binary logistic regression analysis showed that female was an independent risk factor for sarcopenia in MM patients. The survival rates at 2, 3, 4, and 5 years were 87.9%, 71.8%, 64.4%, and 53.7% in the sarcopenia group, and 92.1%, 75.8%, 66.8%, and 66.8% in the normal muscle group, respectively, with no statistically significant differences ( HR=0.71, P=0.364). Conclusion:The incidence of radiation-based sarcopenia in MM patients is 53.0%. Low hemoglobin and blood creatinine levels, not receiving stem cell transplantation, and recurrence or progression of MM are associated with sarcopenia in MM patients, and female is an independent risk factor for sarcopenia in MM patients.