1.Study of the Relationship between Angiogenic Factors and Mechanism of Preeclampsia
Jiwen ZHU ; Xiumin NIU ; Lin ZHAO ; Huabing LI
Tianjin Medical Journal 2009;37(12):1017-1019
Objective:To study the relationship between serum soluble intercellular adhesion molecule-1(sICAM-1) and vascular endothelial growth factor(VEGF), placenta growth factor(PLGF), vascular endothelial growth factor receptor 1(VEGFR1, Flt-1), soluble vascular endothelial growth factor receptor 1(sVEGFR1, sFlt-1) mRNA expression in placenta tissue of preeclampsia(PE). Methods: The serum level of sICAM-1 was measured by enzyme-linked immunosorbent assay and gene expression of placenta tissue was detected by quantitative reverse transcriptase-polymerase chain reaction(RT-PCR). Results:(1)The average serum level of sICAM-1 was(218.45±29.93) μg/L in PE group compared with (168.84±19.39) μg/L in controls(P < 0.01).(2)The mRNA expressions of VEGF, PLGF, Flt-1 and sFlt-1 were increased in placenta of PE than those in controls (P < 0.01).(3)There was a positive correlation between the serum level of sICAM-1 and the sFlt-1 mRNA expression in placenta tissue(r = 0.90, P < 0.01). Conclusion: The serum level of sICAM was increased in the patients with PE. The expressions of VEGF, PLGF, Flt-1 and sFlt-1mRNA were increased remarkably in the placenta tissue of palients with PE,especial for sFlt-1. The remarkable increase for expression of sFlt-1mRNA may contribute to endothelial dysfunction, hypertension and proteinuria.
2.Study of Bilingual teaching in Neurology for Seven-year Medical Master
Lijie CHEN ; Bingrong LIU ; Weizhi WANG ; Yulan ZHU ; Huabing WANG
Chinese Journal of Medical Education Research 2003;0(04):-
We investigated the students of a seven-year medical master in the bilingual teaching program in Neurology with a questionnaire.We found that the bilingual teaching program has the following problems:the students don't have a solid foundation of medical English terminology and the content of the English teaching was higher than the ability of the students.So we think that we need to impress upon the students the importance of the program.We can also raise the overall quality of this program through an improvement of teaching methods,regulation of the proportion of English content in all courses and suitable midterm and final examinations.These alterations will effectively improve the bilingual teaching program.
3.Immune responses in mice induced by DNA vaccines containing different glycoprotein C (Gc) gene fragments of Xinjiang hemorrhagic fever virus
Meifang WANG ; Chaofan GUO ; Huabing ZHU ; Lijuan CHEN ; Yujiang ZHANG ; Surong SUN
Chinese Journal of Microbiology and Immunology 2017;37(6):443-448
Objective To construct two DNA vaccines based on two glycoprotein antigen segments of Xinjiang hemorrhagic fever virus (XHFV) and to evaluate the immune responses in BALB/c mice following vaccination.Methods Two recombinant expression plasmids pVAX1-GcⅠand pVAX1-Gc Ⅱ were constructed by inserting XHFV YL04057 strain Gc Ⅰ (1 229-1 349 aa) and Gc Ⅱ (1 443-1 566 aa) fragments into the eukaryotic expression vector pVAX1 and then were identify by restriction enzyme digestion and sequencing analysis.The recombinant expression plasmids were transfected into mice by hydrodynamics-based transfection.Immune responses induced in mice were evaluated by testing the proliferation of T cells with MTT,measuring serum antibody level with ELISA and detecting cytokines in the supernatant of spleen cell culture with ELISA kit.Results The recombinant expression plasmids were successfully constructed as indicated by the results of restriction enzyme digestion and sequencing analysis.Expression of Gc Ⅰ and Gc Ⅱ genes in mice liver tissues was detected.Antibody titers in mice immunized with pVAX1-GcⅠor pVAX1-Gc Ⅱ were higher than those in mice immunized with pVAX1.Compared with pVAX1,pVAX1-Gc Ⅱ significantly enhanced the proliferation of splenic T lymphocytes and the expression of IFN-γ (P<0.01).Conclusion The constructed two DNA vaccines for XHFV can induce specific humoral and cellular immune responses in mice.pVAX1-Gc Ⅱ is better than pVAX1-GcⅠin immunogenicity and protective efficacy,suggesting that it can be used as a promising candidate for the development of DNA vaccine for XHFV.
4.Antigenicity and immunogenicity analysis of Xinjiang hemorrhagic fever virus Gc antigen fragment
Jingyuan ZHANG ; Meifang WANG ; Chaofan GUO ; Huabing ZHU ; Yijie LI ; Yujiang ZHANG ; Surong SUN
International Journal of Biomedical Engineering 2019;42(3):185-192,204
Objective To express and purify two domains GcⅠand GcⅡof Xinjiang hemorrhagic fever virus (XHFV) glycoprotein, and to study its immunogenicity and the effects on immune response in mice. Methods The prokaryotic expression plasmids of pET28a-GcⅠand pET32a-GcⅡwere constructed and transformed into E. coli BL21, respectively. The expression and purification conditions of rGcⅠand rGcⅡproteins were optimized. The antigenicity of the fusion protein was detected by Western Blot and enzyme-linked immunosorbent assay (ELISA). BALB/c mice were immunized by protein immunization and DNA priming-protein boosting. The mice were randomly divided into 5 groups, including pVAX1-GcⅠ+rGcⅠgroup, pVAX1-GcⅡ+rGcⅡgroup, rGcⅠgroup, rGcⅡgroup and saline group (control group) with 7 mice in each group. The serum antibody titer of mice was detected by indirect ELISA, and the immune effect was evaluated by spleen T lymphocyte proliferation assay and cytokine content determination. Results The fusion proteins rGcⅠand rGcⅡwere purified and obtained, which could react with positive serum of sheep and had good antigenicity. After three immunizations, the IgG levels in the serum of each experimental group were significantly higher than those in the control group (all P<0.001). The serum antibody titers of the experimental groups were reached above 1:12800. Among them, the concentration of Th2 type cytokine interleukin-4 (IL-4) in the spleen cell culture supernatant of rGcⅡ[(79.97±7.47) ng/L] and pVAX1-GcⅡ+rGcⅡgroup [(61.43±9.27) ng/L] was significantly higher than (24.29±3.81) ng/L of the control group, respectively (all P<0.01). The highest mass concentration [(42.46 ±2.60) ng/L] of Th1 type cytokine interferon-γ(IFN-γ) was observed in the pVAX1-GcⅡ+rGcⅡ group, which was significantly higher than (20.33±1.67) ng/L of the control group, and the difference was statistically significant (P<0.001). That showed a significant antigen-specific splenic T lymphocyte proliferation (P<0.001). Conclusions The purified recombinant proteins rGcⅠand rGcⅡhave good immunogenicity, which can make the immune system T lymphocytes tend to Th2 response, and pVAX1-GcⅡ combined with recombinant protein GcⅡ can induce better antigen-specific immune effect. And pVAX1-GcⅡ combined with recombinant protein GcⅡis expected to be used as vaccine candidates for the prevention and control of XHFV.
5.Syndrome of Inappropriate Antidiuresis-report of 40 cases
Lingling XU ; Feng GU ; Mei LI ; Yong FU ; Huabing ZHANG ; Wei LI ; Lin LU ; Xin YUE ; Mingming HU ; Yan JIANG ; Huijuan ZHU ; Ou WANG ; Naishi LI ; Jiangfeng MAO ; Jianping XU ; Lian DUAN ; Shi CHEN ; Ting ZHOU
Chinese Journal of Practical Internal Medicine 2000;0(11):-
Objective We have summarized the clinical characteristics of inappropriate antidiuresis(SIAD).Methods We adopted retrospective analysis to analyze the clinical and lab data of 40 cases.Results The most common causes of SIAD were malignant tumor,lung disease,and central nervous system disease.The five major abnormal lab data were:hypochloraemia,hypouricemia,hyponitremia,hypocalcemia,and low hematocrit.Conclusion It is important to diagnose SIAD as soon as possible,and patient presented hyponatremia combined with hypouricemia must be suspected to have SIAD.
6.Isolation,culture and identification of human skin epidermal stem cellexosomes
Biyou Li ; Jie Ma ; Qiyu Zhang ; Huabing Zhang ; Yunping Zhu
Acta Universitatis Medicinalis Anhui 2023;58(2):224-229
Objective:
To explore method for isolating and culturing human epidermal stem cells ( EPSCs) in vitro and isolating and purifying epidermal stem cell exsomes ( EPSCs-Exo) by optimizing the technical process.
Method:
Firstly,the improved separating enzyme was used to isolate the EPSCs derived from human skin tissue.Then,an improved serum-free culture medium and 10 specific factors were combined to construct optimized 2D culture medium which could stimulate the growth of EPSCs,promote the secretion of EPSCs-Exo,maintain the stemness and proliferation of EPSCs,and delay the differentiation and maturation of EPSCs. Further,the conditions of differential centrifugation was optimized,and then the human EPSCs-Exo was successfully extracted with high efficiency and high purity.
Results:
The human skin tissue was confirmed with the expressions of markers for epidermal cells. EPSCs were verified with high expression levels of integrin-α6,integrin-β1,P63 and CK19 by immunofluorescence staining and Western blot. The nanoparticle tracking analysis results showed the particles separated for
EPSCs supernatant was saucepan with the detected diameter between 30 - 150 nm. The Western blot results showed the positive expression of membrane markers Tsg101,CD9 and CD63 and the negative expression of intracellular markers Calnexin and GAPDH.
Conclusion
The results show that the human-derived EPSCs have been successfully isolated and cultured in vitro,and the EPSCs-Exo have been successfully isolated and identified.