1.Construction of eukaryotic expression plasmid of Smad-7 and study on its function as TGF-β1 inhibitor
Ping ZHOU ; Jie LI ; Jianzhen FANG
Journal of Chinese Physician 2008;10(7):873-875
Objective To construct eukaryctie expression vectors stably expressing Smad-7 and study its function as TGF-β1 inhibi- tor. Methods Smad-7 genes were obtained by PCR from fetal liver cDNA library and inserted in to eukaryotic expression plasmid pcD- NA3. 1/myc-his-B (-). The constructed plasmids containing right sequence of target genes were transfected into human mesangial cells by lipofectamine. The ceils were selected by G418, the expression of Smad-7 was detected by Western-blotting. Cell proliferation was detected by MTT method, and cell cycle was detected by Flow Cytometer to study its function as TGF-β1 inhibitor. Results We successfully con- struct Smad-7 expression plesmid. Smad-7 can inhibit the effects of TGF-[M induced GI cell cycle block and apoptosis. Conclusion The eukaryotic expression vector of Smad-7 has been successfully constructed. Over-expression of exogenous Smad-7 can inhibit the effects of TGF-β1 induced GI cell cycle block and apoptosis.
2.Outcome analysis of invasive fungal infections in patients with hematologic malignancy after micafungin treatment
Bingxin ZHOU ; Jianzhen SHEN ; Yanjuan LIN
Chinese Journal of Practical Internal Medicine 2006;0(24):-
Objective To investigate the efficacy of micafungin sodium for injection in the treatment of invasive fungal infections in patients with hematologic malignancy.Methods The therapeutic effect,action time and adverse effect were analyzed in 12 cases of invasive fungal infections in patients with hematologic malignancy who underwent intravenous injection of micafungin sodium 100 mg qd in Research Center of Hemotology,Union Hospital affiliated to Fujian Medical University,from February 2007 to October 2007.Results The total effective rate was 66.7%.The effective rates of the patients with clinical diagnosis and with recommended diagnosis were 57.14% and 80% respectively ;there was no significant difference between them.The mean action time was 1~5 days.Conclusion Micafungin sodium for injection is effective and safe in treatment of invasive fungal infections in patients with hematological malignancy.
3.Expression of human beta-defensins in ocular surface tissue
Zhensheng GU ; Yaohua SHENG ; Zhaorong ZHU ; Jianzhen ZHOU ; Shanglin LU
Chinese Journal of Tissue Engineering Research 2009;13(50):9826-9831
BACKGROUND: Human β-defensin is mainly located in various tissues epidermis or epithelium, also exists in ocular surface, but its ocular surface and its role of ocular surface diseases remain poorly understood.OBJECTIVE: To observe the distribution of human β-defensins in ocular surface tissue, and to analyze their potential effects on ocular surface disease. DESIGN, TIME AND SETTING: In vitro controlled observation with regard to ocular surface tissue was performed at the Central Laboratory, Xinhua Hospital, Medical College of Shanghai Jiao Tong University and Cell Biochemistry Institute, Chinese Academy of Sciences Shanghai Branch, between October 2006 and December 2007.MATERIALS: A total of 18 inflammatory conjunctival specimens consisted of 6 pterygium surface bulbar conjunctiva, 4 bulbar conjunctival cysts, 4 acid burn conjunctiva, 2 thermal burn conjunctiva and 2 conjunctival granuloma; 15 inflammatory corneal specimens included 6 viral keratitis, 4 fungal keratitis, 3 bacterial keratitis and 2 eye removal following corneal perforation; 9 cadaver normal bulbar conjunctiva samples, 8 cadaver normal corneal samples. METHODS: RT-PCR method and immunohistochemistry were applied to detect human β-defensin expression in 50 samples. MAIN OUTCOME MEASURES: Distribution and location of human β-defensin proteins in normal and inflammatory ocular surface tissues. RESULTS: RT-PCR showed that human β-defensin 1 and human β-defensin 3 were positive in all of the tested samples, whereas human β-defensin 2 existed in a majority of inflammatory ocular surface tissues and no expression was observed in normal ocular surface tissues. Immunohistochemistry analysis revealed most of inflammatory ocular surface tissues expressed human β-defensins 1 and 2, distributing in epithelial cell layer and predominantly in basal lamina, occasionally infiltration of stromal cells was observed, only a small number of human β-defensin 2 expression was absent; normal cornea and conjunctiva samples presented with human β-defensin 1 expression, distributing in epithelial cells and predominantly in basal lamina, only few expressed human β-defensin 2.CONCLUSION: Human β-defensin 1 and 3 appear to be constitutively expressed in surface epithelial cells and basal lamina of normal and inflammatory ocular surface tissues, while human β-defensin 2 may be induced to express in the majority of inflammatory ocular surface tissues. Three human β-defensins expression plays a pivotal role in preventing ocular surface infection and promoting ocular surface injury repair.
4.The establishment and evaluation of diagnostic accuracy of AllGlo~(TM) probe-based techniques for invasive aspergillosis
Dansen WU ; Jianzhen SHEN ; Xiaoqiang ZHOU ; Songfei SHEN ; Xuemei WU
Chinese Journal of Internal Medicine 2010;49(2):142-145
Objective To establish and evaluate a diagnostic technique based on the AllGlo~(TM) probe for the invasive aspergillosis. Methods With the self-designed AllGlo~(TM) probes and primers and the standards, two standard curves of the real-time PCR based on AllGlo~(TM) probes were established respectively for A. flaws and A. fumigatus,then its specificity, sensitivity and reproducibility were evaluated respectively. Results The findings indicated that the standard curve of A. flavus was Y = - 3. 003X + 36. 825, and A. fumigatus' was Y = - 3. 052X + 38.016, and their interassay coefficient of variation respectively were 15.60% and 12. 94% , suggesting a good reproducibility. The lowest spore concentration they could be detected was 10 CFU/ml, which equated to 100-1000 copies of internal transcribed spacer (ITS)2 genes, suggesting a good sensibility. They didn't have cross-positive reaction with other fungus, human genome and bacteria, which indicated a good specificity. Conclusion The diagnostic technique based on the AllGlo?probe for the invasive aspergillosis possessed a good sensitivity, good specificity and deadly accuracy.
5.A retrospective analysis of 7 cases with Brucellosis in Fujian Province
Feng ZHANG ; Jianzhen SHEN ; Haiying FU ; Huarong ZHOU
Chinese Journal of Endemiology 2016;35(10):768-771
Objective To investigate the clinical features of patients with brucellosis in Fujian Province and to update the relevant physician's knowledge on brucellosis.Method Retrospective analysis of the epidemiological,clinical,laboratory,imaging,treatment and outcome data of 7 patients with brucellosis were carried out in Fujian Medical University Union Hospital between January 2011 and July 2015.Results Of the 7 patients with brucellosis,6 were males and 1 was female aged between 29 to 61 years old,and 6 cases had a history of contact with goat.The main clinical presentations were repeated limb pain in 6 cases,6 patients got fever,4 cases with hepatomegaly,6 cases splenomegaly,and 7 cases lung inflammation.Hematology examination results indicated that 4 cases were normal,1 case decreased and 2 cases increased in white blood cells.Six cases had a liver dysfunction and 4 cases with low albumin level.All patients were detected positive of Malta Brucella by blood culture or bone marrow culture.In 6 cases and 4 cases out of the 6 cases,splenomegaly and hepatomegaly were found through B ultrasonography,respectively.After treatment of the 7 cases by doxycycline combined with minocycline rifampicin,3 cases were recovered,2 cases were relapsed,and 2 cases were improved.Conclusions The clinical symptoms are typical in these brucellosis patients.What we need to do is focus on improving the clinical and related physician's awareness of brucellosis,collecting a comprehensive history,choosing suitable assistant examinations based on the conditions of patients,strengthening the contact between clinical departments and auxiliary departments,and timely diagnosis.
6.Detection of p15 methylation and deletion status in acute lymphoblastic leukemia using hn-MSP
Fuan LIN ; Baoguo YE ; Jianzhen SHEN ; Congmeng LIN ; Liping FAN ; Huarong ZHOU ; Haiying FU
Journal of Leukemia & Lymphoma 2012;21(4):208-212
Objective To explore the characteristics of hemi-nested methylation specific polymerase chain reaction (hn-MSP) and to find out the possible relationship between patterns of methylation or deletion and the developmet of adult acute lymphoblastic leukemia(ALL).Methods hn-MSP and bisulfit-sequencing PCR (BSP) were designed and adopted to analyze p15 gene methylation or deletion patterns in 25 adult ALL patients,malignant hematopathy cell lines and normal lymphocytes. hn-MSP and BSP products were cloned and sequenced.The sensitivity and specificity of hn-MSP were also analized.Results The sequencing results of hn-MSP and BSP products were consistent, and the sensitivity of detection of p15 methylation was up to 1.0×10-5.17 adult ALL patients (68 %) were p15 gene hypermethylation and 3 patients were with deletion of p15 gene exon 1.There were no hypermethylation or deletion in the 10 controls.Conclusions The detection rate of p15 methylation in many tumors,especially in adult ALL,is frequent high.hn-MSP is highly sensitive and specific in analyzing p15 methylation.
7.Qualitative research on the non-compliance of fluid intake among maintence hemodialysis patients
Jianzhen FAN ; Aiyan DU ; Renyan XUN ; Yunjuan HUANG ; Tingli ZHU ; Wei ZHOU
Modern Clinical Nursing 2017;16(2):38-42
Objective To explore the results of non-compliance of fluid intake in maintence hemodialysis patients.Methods The qualitative phenomenological research method was adopted in this study.Self-structured in-depth interviews were conducted with 13 maintenance hemodialysis patients.Data were analyzed by content analysis.Result Four themes were extracted including allotrigeusia,have a smattering of fluid intake knowledge,self-condemned and guilty,muddling along.Conclusions Fuild restriction is the most difficuilt prescribed treatment schedule among maintence hemodialysis patients.Nurses should pay attention to those people and provide multilevel,continual,individual and comprehensive measures.
8.Promoter methylation status of SFRP genes and induced apoptosis by demethylation in Jurkat cells.
Chengbo XU ; Jianzhen SHEN ; Bin LIAO ; Haiying FU ; Huarong ZHOU ; Yan QI ; Zhenping HUANGFU ; Yining CHEN ; Jiawei CHEN
Chinese Journal of Hematology 2016;37(1):51-55
OBJECTIVETo study the promoter methylation status of SFRP genes and the effect of 5- aza- 2'- deoxycytidine (5- Aza- CdR)induced apoptosis via Wnt/β- catenin pathway by demethylation in Jurkat cells.
METHODSJurkat cells were treated with different concentrations of 5- Aza- CdR. The cell proliferation level of Jurkat cells was detected by MTT assay. Apoptosis was evaluated by flow cytometry. Methylation- spcific PCR (MSP) was used to determine the methylation status of SFRP genes. The expressions of SFRP genes were detected by real time fluorescence quantitative PCR. The mRNA expression levels of survivin, c- myc and cyclin- D1 were analyzed by RT- PCR. Western blot was used to detect the levels of β-catenin protein.
RESULTSCompared with control group, the different concentrations of 5-Aza-CdR could significantly inhibit the proliferation of Jurkat cells in a time-dose dependent manner (P<0.05). After being treated by 5- Aza- CdR for 48 hours, the cell early apoptosis rate in experiment group was significantly higher than that in control group (P<0.05). The promoters of SFRP1, SFRP2, SFRP4, SFRP5 genes were hypermethylation state in the control group, after being treated by 5-Aza-CdR for 72 hours, the brightness of SFRP1, SFRP2, SFRP4, SFRP5 genes' methylation strips weakened in a dose- dependent manner. SFRP mRNA expression increased (P<0.05) when 5- Aza- CdR concentration increased, and the level of β- catenin protein was dampened in a dose- dependent manner (P<0.05). As compared to the control group, the mRNA expressions of associated apoptosis genes survivin, c-myc and cyclin- D1, respectively were obviously down- regulated in a dose- dependent manner (P<0.05).
CONCLUSIONThe effect of demethylation could up- regulate SFRP genes expressions by reversing its hypermethylation and induced apoptosis by down-regulation of β-catenin and associated apoptosis genes.
Apoptosis ; Azacitidine ; analogs & derivatives ; pharmacology ; Cell Proliferation ; DNA Methylation ; Down-Regulation ; Gene Expression ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; Jurkat Cells ; Membrane Proteins ; genetics ; Promoter Regions, Genetic ; Wnt Signaling Pathway ; beta Catenin ; metabolism
10.Primary ocular adnexal double-expression diffuse large B-cell lymphoma treated with R-MAD regimen: report of one case and review of literature
Yanquan LIU ; Jianzhen SHEN ; Haiying FU ; Huarong ZHOU ; Langhui ZHANG ; Weili ZHENG
Journal of Leukemia & Lymphoma 2020;29(4):240-243
Objective:To explore the clinical effect of R-MAD (rituximab, methotrexate, cytarabine, dexamethasone) regimen in the treatment of primary ocular adnexal double-expression diffuse large B-cell lymphoma (DLBCL).Methods:The clinical data of an elderly patient with primary ocular adnexal double-expression DLBCL who was treated with R-MAD regimen in June 2019 in Fujian Medical University Union Hospital was retrospectively analyzed. The clinical manifestations, diagnosis and treatment, prognosis were also analyzed, and the related literature was reviewed.Results:The patients was a 71-year-old male. After initial treatment of R-CHOP chemotherapy, the patient's eye mass did not shrink, the swelling and pain became worse, the curative effect was not good, and the disease progression continued. After the patient was given R-MAD chemotherapy for 3 courses, the eye swelling subsided and pain symptoms were significantly improved, satisfactory results were obtained, and no obvious adverse reactions occurred.Conclusions:R-MAD regimen has an ideal effect on the patient with primary ocular adnexal double-expression DLBCL, which can significantly improve symptoms, delay disease progression, and improve the quality of life of patients, but the prognosis still needs to be followed up in the long-term.