1.Cloning,Subcellular Localization and Expression Analysis of AaCPS from Artemisia argyi
Xiang CHENG ; Yue MING ; Jingmao YOU ; Zhaohua SHI ; Bo WANG ; Dingrong WAN ; Xianming LIN ; Ran XU
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(5):1223-1232
Objective To clone the full-length sequence,subcellular localization,and analyze the expression patterns of the β-caryophyllene synthase gene(AaCPS)of Artemisia argyi,in order to lay a foundation for the gene function analysis of sesquiterpene biosynthesis pathway in A.argyi.Methods The genes annotated as CPS were selected from the transcriptome data of A.argyi.The full-length cDNA sequence of the target gene was obtained by PCR,and the coding region was analyzed using bioinformatics.A prokaryotic expression vector was constructed and transformed into Escherichia coli competent cells to express recombinant protein.A green fluorescent protein(GFP)fused expression vector was constructed,and the subcellular localization of AaCPS was observed using Agrobacterium tumefaciens transient expression method in tobacco.Real-time quantitative PCR(qRT-PCR)was used to analyze the expression patterns of the AaCPS at different harvest times(April,May,June,July).Determination of β-caryophyllene by HS-SPME-GC-MS and correlation analysis.Results The AaCPS gene was cloned from A.argyi.The full-length open reading frame(ORF)was 1647 bp,encoding 548 amino acids.The molecular weight was 63 667 Da with a theoretical pI of 5.94.AaCPS contained conserved Terpen_synth and Terpen_synth_C domains.Phylogenetic analysis indicated that AaCPS was closely related to the CPS protein of Artemisia annua.SDS-PAGE showed the presence of target protein bands between 75 000-120 000 Da(containing 28 132 Da of the labeled protein),indicating successful expression of the AaCPS protein.The protein was found to be localized in the cytoplasm.As shown by qRT-PCR,the expression of AaCPS gene showed an upward trend,reaching the highest in July.The results of HS-SPME-GC-MS showed that the content of β-caryophyllene increased gradually from April to the highest in June.It is consistent with the trend of AaCPS gene expression from April to June.Conclusion Through the cloning and analysis of AaCPS gene,a foundation has been laid for further study of the functional identification of the AaCPS gene in the sesquiterpenes biosynthesis pathway in A.argyi.
2.Correlation analysis of serum zonulin, ZO-1, TNF-α with severity of acute pancreatitis
Shuaiting MA ; Ming GAO ; He LI ; Chunlin YIN ; Haiping WANG ; Jun CHENG ; Zhaohua WANG ; Kai SONG ; Yuansong SUN
Journal of Chinese Physician 2023;25(7):1041-1045
Objective:To analyze the correlation between the severity of acute pancreatitis (AP) and the levels of zonulin, zonula occludens protein-1 (ZO-1), tumor necrosis factor -α (TNF -α) in the peripheral blood of patients with acute pancreatitis (AP), and the value of predicting moderate and severe AP.Methods:The clinical data of 115 AP patients admitted to the Second Affiliated Hospital of Anhui Medical University from June 2020 to January 2022 were retrospectively analyzed. They were divided into mild group (69 cases) and moderate severe group (46 cases). The blood levels of zonulin, ZO-1, and TNF-α were measured for all patients on the 1st, 3rd, and 7th day after admission, and the results of the two group tests were compared. The correlation between zonulin, ZO-1, TNF -α and Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) scores on the 1st day was and the value of various indicators for predicting moderate to severe AP were analyzed.Results:The C-reactive protein (CRP) levels of AP patients in the moderate to severe group were higher than those in the mild group, and the difference was statistically significant (all P<0.05). The levels of zonulin, ZO-1, and TNF -α in AP patients in the moderate to severe group showed an upward trend on the 1st, 3rd, and 7th days after admission. The levels of zonulin, ZO-1, and TNF -α in AP patients in the moderate to severe group were higher than those in the mild group at the same time point, and the differences were statistically significant (all P<0.05). The APACHE Ⅱ score of AP patients on the first day of admission was positively correlated with the levels of zonulin, ZO-1, and TNF -α ( r=0.736, 0.552, 0.621, all P<0.05). Zonulin had the highest area under the curve (AUC) for predicting moderate to severe AP, at 0.892, with an optimal threshold of 2.075 pg/ml. Zonulin had the highest sensitivity, at 0.804, and ZO-1 had the highest specificity, at 0.926. Using zonulin ≥2.075 pg/ml, ZO-1≥399.4 ng/ml, and TNF -α≥40.88 pg/ml as thresholds; the sensitivity and specificity obtained from parallel experiments were 0.976 and 0.710, respectively; The sensitivity and specificity obtained from the series of experiments were 0.326 and 0.999, respectively. Conclusions:There is a correlation between the serum levels of zonulin, ZO-1, and TNF -α in AP patients and the severity of AP. Zonulin, ZO-1, and TNF -α have certain clinical value in predicting moderate to severe AP.
3.Astragaloside in cancer chemoprevention and therapy.
Yaning WANG ; Zhuo ZHANG ; Zhaohua CHENG ; Wei XIE ; Hanjiao QIN ; Jiyao SHENG
Chinese Medical Journal 2023;136(10):1144-1154
Tumor chemoprevention and treatment are two approaches aimed at improving the survival of patients with cancers. An ideal anti-tumor drug is that which not only kills tumor cells but also alleviates tumor-causing risk factors, such as precancerous lesions, and prevents tumor recurrence. Chinese herbal monomers are considered to be ideal treatment agents due to their multi-target effects. Astragaloside has been shown to possess tumor chemoprevention, direct anti-tumor, and chemotherapeutic drug sensitization effects. In this paper, we review the effects of astragaloside on tumor prevention and treatment and provide directions for further research.
Humans
;
Chemoprevention
;
Antineoplastic Agents
;
Neoplasms/prevention & control*
;
Saponins/pharmacology*
;
Triterpenes/pharmacology*
4.Qualitative study of disease cognition in patients with recurrent acute pancreatitis
Yanjie GUO ; Xue DONG ; Zhaohua CHENG ; Xuemei ZHANG ; Mingming ZHANG
Chinese Journal of Modern Nursing 2023;29(15):2039-2044
Objective:To understand the experience of inpatients with recurrent acute pancreatitis and to provide empirical support for health intervention and health education for medical staff.Methods:Using the descriptive phenomenology, a total of 11 inpatients with recurrent acute pancreatitis from the Second Norman Bethune Hospital of Jilin University were selected for semi-structured interviews from October to December 2021 by the objective sampling and theoretical sampling methods. Colaizzi phenomenological analysis method and Nvivo11 coding software were used to analyze and sort out the collected interview data and extract classified topics.Results:Three themes were summarized, including lack of knowledge of patients in acquiring disease knowledge, difficulty in self-control of disease progression under the influence of subjective or objective factors, active or forced changes in dietary behavior and attitudes.Conclusions:Medical staff should strengthen the guidance of acquiring health knowledge of recurrent acute pancreatitis, strive to enhance patients' ability of disease self-management and fully evaluate and standardize the diet of patients with recurrent acute pancreatitis.
5.Research progress of the construction of humanity nursing curriculum system
Yuanjuan CHENG ; Kun LI ; Zhaohua CHENG ; Lili LI ; Yuanyuan SHI ; Yixiu BAI ; Yufei WANG
Chinese Journal of Modern Nursing 2019;25(23):3027-3032
With the transformation of medical model, nursing humanities has gradually become the core of nursing practice, and the reform and construction of nursing humanities curriculum system has become one of the urgent problems to be solved in the reform of nursing education. This paper summarized the research progress of the construction of nursing humanities curriculum system at home and abroad, including curriculum setting, teaching strategies and methods, assessment forms, teacher competence and so on. It is hoped that it will be of reference value to the future construction of medical-teaching synergy of nursing humanities curriculum and the integration model of colleges and universities.
6.Effects of SDF-1/CXCR4 signal pathway blockade by AMD3100 on the adhesion of leukemia cells to osteoblast niche and the drug resistance of leukemia cells.
Zhaohua SHEN ; Dongfeng ZENG ; Zhongmin ZOU ; Lidan ZHU ; Ying MA ; Xi ZHANG ; Cheng ZHANG ; Jiali LI ; Peiyan KONG
Chinese Journal of Hematology 2015;36(5):413-417
OBJECTIVETo study the blocking effect of CXCR4 inhibitor AMD3100 on the adhesion of leukemia cells to osteoblast niche, and the reversal of multidrug resistance in leukemia cells.
METHODSMesenchymal stem cells (MSCs) from leukemia patients were planted on the bio-derived bone scaffolds and then induced into osteoblasts to establish the bio-osteoblast niche. The levels of SDF-1were tested with ELISA. The leukemia cell line MV4-11 cells with FLT3-ITD mutation were inoculated into the bio-osteoblast niche to build a three-dimensional co- culture system. The expression level of CXCR4, adhesion and apoptosis rates of leukemia cells were observed by flow cytometry after incubation with AMD3100 and Ara-C for 24 h and 48 h.
RESULTS(1)The supernatant levels of SDF-1 in cultured osteoblast were (304 ± 18), (410 ± 28) and (396 ± 16) pg/ml on 7 th, 14 th and 21 th day, respectively. It reached the highest on 14 th day. The expression level of CXCR4 in cultured MV4-11 cells was (72 ± 16)%. (2)Adhesion rate of MV4-11 cells to osteoblast niche was (40.1 ± 8.1)% after AMD3100 treatment for 24 h, while that of control group was (65.6 ± 12.1)% (P<0.05). (3)The apoptosis rate of MV4-11 cells incubated with AMD3100 for 24 h was (5.6 ± 0.8)%, while that of control group was (2.5 ± 0.5)%. The apoptosis rates of AMD3100-induced MV4-11 cells were (10.0 ± 2.4)%, (17.8 ± 2.3)% and (25.1 ± 2.4)% after treatment with Ara-C at 0.02, 0.20, 2.00 mg/ml respectively and they were (6.7 ± 1.0)%, (10.3 ± 1.5)%, (16.2 ± 3.1)% respectively in AMD3100-noninduced control group, the difference was significant (P<0.05).
CONCLUSIONAMD3100 can block the interaction between osteoblasts niches and leukemia cells, and play an important role in the reversal of multidrug resistance in leukemia cells.
Apoptosis ; Cell Adhesion ; Cell Line, Tumor ; Chemokine CXCL12 ; Coculture Techniques ; Cytarabine ; Drug Resistance ; Flow Cytometry ; Heterocyclic Compounds ; Humans ; Leukemia ; Mesenchymal Stromal Cells ; Osteoblasts ; Receptors, CXCR4 ; Signal Transduction
7.Clinical efficacy observation of stereotactic radiation therapy combined with gemcitabine for advanced pancreatic carcinoma
Jianping DAI ; Fan WANG ; Hegao WANG ; Ruisong GUO ; Zhaohua LIU ; Wenhua CHENG ; Xiaojuan LIU
Cancer Research and Clinic 2014;26(1):45-47
Objective To seek the curative effect of stereotactic radiation therapy (SRT) combined with gemcitabine for unresectable advanced pancreatic carcinoma.Methods 24 patients were treated by SRT of 6MV X-ray.Patients were fixed with the rack of stereotactic localization and heated plastics mould.The CT scanning results were put into the treatment planning system.According to the target area of tumor,sensitive organs and moving error drew GTV,CTV and PTV.The best plan was selected by the dose-volume histogram (DVH).5 to 7 beams of non-coplanar radiation ray were chosen.PTV was surrounded by ≥95 % isodose curves.Conventional fraction,5 fractions per week,1.8-2 Gy per fraction was used.All patients received a total dose of 50-60 Gy.Gemcitabine was performed 1000 mg/m2,once a week,iv gtt,in the 1st,2nd and 4th,5 th weeks.Results In chemo-radiotherapy,50.0 % (12/24) patients showed light nausea,41.7 % (10/24) patients showed leucopenia or thrombocytopenia of grade 1 or 2,after symptomatic treatment,all patients completed the planned treatment.In a period of one month to three months,after SRT combined with chemotherapy,appetite improvement was 83.3 % (20/24),jaundice disappeared in 6 of 6 patients (100.0 %),abdominal pain was relieved in 21 of 24 patients (87.5 %),3 patients were relieved completely among them.The complete remission (CR) rate was 16.7 % (4/24) and partial remission (PR) rate was 66.7 % (16/24),with CR+PR rate of 83.4 % (20/24).One-year survival rate was 70.8 % (17/24).As a consequence of cachexia,intestinal obstruction or bleeding,7 patients died within one year.Nobody survived more than 2 years.Conclusions SRT combined with gemcitabine for advanced pancreatic carcinoma may relieve symptoms.It is an effective approach to improve life quality and prolong survival time for advanced pancreatic carcinoma,especially lod and weak patients are more suitable to select SRT combined with gemcitabine.
8.Influence of curcumin in NO and S1 00βlevels in brain tissue of rats after focal cerebral ischemia reperfusion inj ury
Zhaohua ZHAO ; Shuqiang WU ; Xingchun GOU ; Yajing MI ; Jiping YANG ; Lili SHI ; Juanjuan CHENG
Journal of Jilin University(Medicine Edition) 2014;(5):925-928
to observe the infarction volume.Nitrate reductase assay was used to detect the level of NO in brain tissue of the rats.The level of S100βin brain was detected by ELISA method.Results Compared with model group,the brain infarction volumes of the rats 24 and 72 h after cerebral ischemia reperfusion in curcumin group were significantly decreased (P<0.05).Compared with sham operation group,the NO and S100βlevels in the brain tissue 24 and 72 h after cerebral ischemia reperfusion of the rats in model group were significantly increased(P<0.05);compared with model group,the levels of NO in the brain tissue 24 and 72 h after cerebral ischemia reperfusion in curcumin group were remarkably decreased (P<0.05);compared with modee group,the level of S100βin the brain tissue 72 h after cerebral iscemia reperfusion in curcumin group was remarkably decreased (P < 0.05 ). Conclusion Curcumin can significantly reduce the degree of ischemia reperfusion injury in the rats and reduce the levels of NO and S100βin brain tissue,which suggests that the decrease of NO and S100βlevels in brain tissue may be associated with the neuroprotective effect of curcumin.
9.Research on effects of bone marrow mononuclear cells implantation on model of experimental pulmonary artery hypertension.
Yan LU ; Zhaohua ZHANG ; Guanghui CHENG ; Yun LUAN
Journal of Biomedical Engineering 2013;30(3):601-606
In the present study, we carried out intratracheal administration of bone marrow-derived mononuclear cells (BM-MNCs) to dehydromonocrotaline (DMCT)-induced canine pulmonary artery hypertension (PH) of rat model to examine the security and feasibility, and the aim was to discuss the mechanism. All animals (n=30) were randomly divided into 3 groups (n=10 in each group), i. e. control group, PH group and BM-MNCs group. Six weeks after the transplantation, the hemodynamic data and right ventricle weight ratio were significantly improved for those in BM-MNCs group compared with those in PH group. The lung mRNA levels of vascular endothelial growth factor (VEGF) were higher, while preproendothelin-1 (ppET-1), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were lower compared with those in the PH group (P<0. 05). Immunofluorescence and histochemical results confirmed that 6 weeks after the administration, transplanted BM-MNCs were still alive and could differentiate into pulmonary vascular endothelial cells. These results showed that intratracheal administration of BM-MNCs could obviously reduce or even reverse the DMCT induction of PAH process. The mechanism could be explained as that the function was mainly through the paracrine effect to promote renewable and reduce inflammation.
Animals
;
Bone Marrow Cells
;
cytology
;
Cell Transplantation
;
methods
;
Dogs
;
Familial Primary Pulmonary Hypertension
;
Female
;
Hypertension, Pulmonary
;
chemically induced
;
therapy
;
Leukocytes, Mononuclear
;
transplantation
;
Male
;
Monocrotaline
;
analogs & derivatives
;
Rats
10.Investigation on clinical efficacy of the fourth-generation ultrasonic lithotripsy system in the treatment of urinary tract stones
Zhaohua LIU ; Zhijian YANG ; Minjian CHEN ; Bingquan WU ; Cheng PANG ; Xihai LIU
Chinese Journal of Primary Medicine and Pharmacy 2012;19(7):978-980
ObjectiveTo evaluate the fourth-generation ultrasonic lithotripsy system for treating the urinary tract stones.Methods243 cases of urinary tract stones who received treatment of fourth-generation EMS ultrasonic lithotripsy system were analyzed retrospectively.ResultsImmediate phase I lithotrotrisy was performed successfully in 227 cases and the successful rate of phase I was 93.3% (227/243).Delayed phase Ⅱ lithotripsy was performed for 16 patients.Complications happened in 2 cases,one case occurred hydropneumothorax during operation,required indwelling thoracic drainage tube processing,the other had severe bleeding which conservative treatment was ineffective,cured by the intervention of super-selective renal artery embolization.Conclusion Fourth generation of ultrasonic lithotripsy for treating stones on different anatomical locations of urinary system was safe,practical and efficient,worthy of clinical application.

Result Analysis
Print
Save
E-mail