1.Effect of Diagnostic Ultrasound on SOD of Mother and Progeny in Early Pregnancy
Lirong HUO ; Ming YAN ; Jinmin LIU
Journal of Medical Research 2006;0(12):-
0.05).②The animal experiment showed ,in contrast to the control , embryonate rats’ rhodocyte SOD were significantly different after 2 days but no difference before prenatal. ③in contrast with the control , SOD of placenta were significantly different only after 9 days. Conclusions Diagnostic ultrasound in early pregnancy can make SOD change ,but the changes can be reparable.
2.Expression of recombinant plasmid of homo heterogeneous nuclear ribonucleoprotein E1 in SH-SY5Y cells
Lirong HUO ; Lanying WANG ; Junhua ZOU ; Nan ZHONG
Chinese Journal of Tissue Engineering Research 2014;(42):6769-6774
BACKGROUND:The functions of homo heterogeneous ribonucleoprotein E1 are very wide. It can participate in the expression of skeleton proteins in the nervous system. OBJECTIVE:To construct the recombinant plasmid of homo heterogeneous ribonucleoprotein E1 and observe its expression in nerve cells for further studying the functions of it in neurocytes. METHODS:Using pcDNATM4/His C, the homo heterogeneous ribonucleoprotein E1 was subcloned into recombinant plasmid E1-pcDNATM 4/His C, fol owed by enzyming and sequencing. After SH-SY5Y cells were transfected with the recombinant plasmid, western blot analysis and real time RT-PCR were used to detect the expression of homo heterogeneous ribonucleoprotein E1 in SH-SY5Y cells. And the growth of SH-SY5Y cells was observed. RESULTS AND CONCLUSION:We successful y constructed the eukaryotic expressed vector of homo heterogeneous ribonucleoprotein E1. The recombinant plasmids were verified to express in SH-SY5Y cells correctly at mRNA and protein levels. And SH-SY5Y cells generated quickly after homo heterogeneous ribonucleoprotein E1 was over-expressed. The homo heterogeneous ribonucleoprotein E1 is an important protein in neural development. And this vector offers the premise for further studying its function in nervous system.
3.Effects of antimicrobial peptide LL-37 expressed and purified from prokaryotes in the murine model of vaginal candidiasis
Fang WANG ; Yan HUO ; Lirong YIN ; Bei SUN ; Pingping ZHANG
Chinese Journal of Obstetrics and Gynecology 2016;51(7):530-534
Objective To study the effects of antimicrobial peptide LL-37 expressed and purified from prokaryotes on candida albicans growth. Methods (1) Thirty female Kunming mice were treated with estrogen and white candida yeast suspension were poured into vagina to establish a vulvovaginal candidiasis (VVC) murine model. After successful establishing the VVC mouse model, mice were randomly sorted into test group (n=15) and control group (n=15) . Suspension(30μl, 100μg/ml)of recombinant peptide LL-37 expressed and purified in Prokaryotes was given by intravaginal administration to the test group for 5 days, while the same amount of phosphate buffered saline (PBS) was given to the control group. (2) Tweenty-four hours after treatment, the fungal burden and colony-forming unit (CFU) of vaginal fluids were evaluated. All mice were subsequently sacrificed and vaginal tissues were harvested for tissue homogenate preparation. ELISA was used to determine the levels of nterleukin-10(IL-10)and interferon-γ(IFN-γ)in the isolated vaginal tissues. Results (1) VVC mouse model was established successfully in this study. Vaginal mucosa congestion, edema, vaginal plica disappearing were obviously observed in the control group. After treatment with recombinant protein LL-37 vaginal mucosa has no obvious change in the test group. (2) Fungal burden and CFU of vaginal fluids were significantly lower in the test group [(4.8±1.0)×104 CFU/ml] than that in the control group [(8.5 ± 2.1) × 104 CFU/ml, P=0.017]. IFN-γlevel of the test group was increased [(257 ± 11) vs (197 ± 4) pg/ml, P=0.000], while the level of IL-10 was reduced [ (287 ± 15) vs (379 ± 17) pg/ml P=0.000] resulting in a the ratio of IFN-γ/IL-10 was in significantly higher in test group (0.892±0.008 vs 0.496±0.013, P=0.000). Conclusion Recombinant protein LL-37 expressed and purified from prokaryotes inhibits the growth candida albicans and improves vaginal immunity by adjusting IFN-γand IL-10 secretion in the VVC mouse model, highlighting the therapeutic potential of LL-37 for VVC.
4.Influence of different administration regimens for treatment of bacterial vaginosis on vaginal microflora and the expression of immune factors
Fang WANG ; Yan HUO ; Yanyan LIU ; Lirong YIN
Tianjin Medical Journal 2016;44(8):1029-1032
Objective Tracking of the vaginal microflora recovery and the expression of immune factors from untreated and treated patients with bacterial vaginosis (BV) by using different administration regimens and studying the relationship of treatment results and regimen selections. Methods 25 healthy females were selected as a control group and 100 BV patients were randomly divided into 4 groups (n=25/group). Group A: Intravaginal administration of metronidazole (× 7 d), Group B:Continuous intravaginal administration of metronidazole (× 7 d) and then live Lactobacillus Capsule (× 7 d) , Group C: Intravaginal administration of nifuratel (× 7 d), Group D: Continuous intravaginal administration of nifuratel (× 7 d) and then live Lactobacillus Capsule (×7 d). The microecological assessment system and EILSA were used to compare the clinical efficacy, vaginal microflora recovery and the changes in IL-8, TLR2 and TNF-αof the vaginal lavage fluid in healthy women or patients with bacterial vaginosis before and after treatments by four treatment strategies. Results ① The vaginal microflora imbalance, flora disturbance, pH value increased were presented in BV group compared with the control group.②Compared to the median of IL-8, TLR2 and TNF-α in vaginal lavage fluids of control group, there was no significant difference in IL-8 level but both TLR2 and TNF-αwere significantly increased (P<0.05) in BV group. The immune factors had no significantly difference in all BV groups.③The therapeutic effect in each BV groups was compared after stopping treatment for 7 days. The cure rate and the vaginal microflora recovery rate were significant higher in group B and D than group A and C (P<0.05). ④ After treatment there was no significant change in IL-8 level but there was an obviouslydecrease in TLR2 and TNF-α(P<0.05). The decreased levels are more significant in groups B and D than groups A and C (P<0.05). Conclusion By combining with the microecological assessment system to evaluate the therapeutic effect of BV, our research suggests that the sequence schemes of nifuratel plus live Lactobacillus Capsule is more effective in therapy effect, restoring normal vaginal micro-ecological environment and vaginal local immunity than metronidazole used alone.
5.Effect on Nervous Tissue and Anaphase Diagnostic Ultrasound in Early Pregnancy.
Lirong HUO ; Jiantao LIANG ; Jinmin LIU ; Wangpeng LIU
Journal of Medical Research 2006;0(02):-
Objective To assess the diagnosis safety by detecting the biological character of prenatal fetus rats’nervous tissue exposed to diagnostic ultrasound during earlier period. Methods Cell culture, morphology examining, cell proliferation curve measurement and flow cytometry detection were adopted. Results ①After morphology examining, there are no difference between the treated group and control group. ② We can see from the growth curve, the two groups has identical growth tendency. ③Through flow cytometry detection, results show that there are no significance between two groups. Conclusion Diagnostic ultrasound in early pregnancy have no significant effect on prenatal fetus rats’nervous tissue.
6.Prokaryotic Expression and Antifungal Activity of HumanαDefensin-5 Protein
Pingping ZHANG ; Lirong YIN ; Fang WANG ; Bei SUN ; Yan HUO
Tianjin Medical Journal 2014;(8):755-758
Objective To construct the prokaryotic expression vector for HD-5 and purify the recombinant HD-5 protein then analyze its antifungal activity. Methods The HD-5 gene was cloned by PCR, then was inserted into prokary-otic expression plasmid pQE-30Xa to construct pQE-30Xa/HD-5. After sequencing, pQE-30Xa/HD-5 was transformed in-to E.coli M15 cells. Its expression was induced by IPTG and confirmed by SDS-PAGE. The recombinant protein was purified through Ni-NTA affinity purification system. The antifungal activity was tested by disk diffusion method. Results HD-5 gene and pQE-30Xa/HD-5 vector were obtained successfully. E.coli M15 strains was used to express HD-5 fusion protein. After purification, the fusion protein was confirmed by Western blot. The disk diffusion test confirmed that the fusion pro-tein can inhibit Candida albicans. Conclusion Expression vector pQE-30Xa/HD-5 was successfully constructed. The HD-5 fusion protein was expressed in E.coli successfully, which showed a certain degree of antifungal activity.
7.Identification of novel partner proteins of PCBP1
Lirong HUO ; Chen SHEN ; Weina JU ; Junhua ZOU ; Wu YAN ; W.ted BROWN ; Nanbert ZHONG
Journal of Peking University(Health Sciences) 2009;41(4):402-408
Objective: PCBP1 is a family member of heterogeneous nuclear ribonucleoproteins (hnRNPs) that belong to RNA-binding proteins and bear three KH domains. The protein plays a pivotal role in post-transcriptional regulation for RNA metabolism and RNA function in gene expression. We hy-pothesized and were going to identify that the regulatory function of PCBP1 is performed through different complexes of proteins that include PCBP1. Methods: To test our hypothesis, approaches of protein wal-king with a yeast two-hybrid system (Y2H), pulling down in yeasts, co-immunoprecipitation and immu-nofluorescent microscopy assay were employed in this study. The PCBP1 was used as the initial "walker" to search for its interaction partner(s). Results: Candidate proteins including MYL6, PECAM1, CSH1,RAB7, p57KIP2, ACTG1, RBMS1 and PSG4-1ike were identified with selection mediums and preceding methods. Conclusion: With these candidate protein molecules, some protein complexes associating with PCBP1 are proposed, which may help in a better understanding of physiological functions of PCBP1 and proved evidence that PCBP1 is involved in variant biological pathways.
8.Prokaryotic expression and purification of antimicrobial peptide LL-37 and the inhibiting effect against Candida albicans
Yan HUO ; Fang WANG ; Bei SUN ; Lirong YIN ; Pingping ZHANG ; Yujuan ZHANG ; Bumei ZHANG
Chinese Journal of Obstetrics and Gynecology 2016;(2):120-125
Objective To study the inhibitory effect of antimicrobial peptide LL-37 on Candida albicans through its ability to promote the secretion of immune factors by vaginal epithelial cells. Methods (1) LL-37 prokaryotic expression vector pET-Duet/LL-37 was constructed and its expression was induced in Escherichia coli M15. The expressed LL-37 fusion protein was purified and identified by western blot. Antifungal activity of the purified protein was initially identified by Kirby-Bauer (K-B) method. (2) Purified LL-37 protein was added to human vaginal epithelial cells co-cultured with Candida, and inhibitory effect on Candida growth was determined by the glucose consumption method. Interferon γ (IFN-γ), interleukin 10 (IL-10) concentration and IFN-γ/IL-10 ratio were measured by ELISA at different time points. Results (1) LL-37 fusion protein was purified to 96% purity at a concentration of 433.92 μg/ml, and was shown to possess anti-fungal activity confirmed by the K-B method. (2) A Candida-vaginal epithelial cells co-culture system was successfully constructed. LL-37 recombinant protein inhibited the growth of Candida with absorbance values significantly higher in the treatment group compared to the control group at all measured time points (12-hour:3.008±0.003 versus 2.967±0.003, 24-hour:2.941±0.003 versus 2.601±0.003, 48-hour:2.893 ± 0.004 versus 2.409 ± 0.003; all P<0.01). Furthermore, the rate of decrease was also much slower compared to the control group. In both control and experimental groups, IFN-γ and IL-10 secretion levels were observed to rise at first peaking at 24 hours and subsequently decrease. For each time period, IFN-γconcentration in the experimental group was significantly higher at 24 hours compared to the control group [(104.00 ± 1.07) versus (85.17 ± 0.28) pg/ml, P<0.01]. In contrast, IL-10 concentrations were significantly lower than the control group at all time points (P<0.01). IFN-γ/IL-10 ratio was also observed to be significantly higher than the control group at all measured time points (P<0.01). Conclusions (1) Recombinant protein LL-37 could significantly inhibit the growth of Candida. (2) By influencing the secretion of immune factors such as IFN-γ, IL-10, etc, recombinant protein LL-37 is able to adjust vaginal epithelial cells local immunity, and enhance resistance to Candida infection.
9.Development and application of ChatGPT in medical research management
Chinese Journal of Medical Science Research Management 2023;36(4):304-307
Objective:To analyze the development prospect of ChatGPT in the field of medical research management and the feasibility of application.Methods:The literature was reviewed and analyzed.Results:This paper describes the application of ChatGPT involving the orderly promotion of medical scientific research projects, the working mode of scientific research managers, and the development of scientific research management platform.Conclusions:The development of ChatGPT makes it have its unique advantages in medical scientific research management, but also has some shortcomings, which need to be reasonably understood and scientifically applied.
10.Earthworm Protein Protects Vascular Endothelial Function in Spontaneously Hypertensive Rats via PI3K/Akt/Nrf2 Signaling Pathway
Huilin ZHAO ; Xiaojing LI ; Lirong WANG ; Yaohui HUO ; Yun LI ; Tianlong DING ; Kai LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(2):118-126
ObjectiveTo observe the effect of earthworm protein on the expression of phosphatidylinositol 3-kinase/protein kinase B/nuclear factor E2-related factor 2 (PI3K/Akt/Nrf2) pathway in the aorta of spontaneously hypertensive rats (SHR) and explore mechanism of earthworm protein in treating hypertensive vascular endothelial dysfunction (VED). MethodTen 10-week-old Wistar Kyoto (WKY) rats and fifty SHR rats were selected for a week of adaptive feeding. WKY rats were selected as the normal group, and fifty SHR rats were randomized according to body weight into model, valsartan (8×10-3 g·kg-1·d-1), and high-, medium-, and low-dose (0.2, 0.1, 0.05 g·kg-1·d-1, respectively) earthworm protein groups. The normal and model groups were administrated with equal volume of double distilled water by gavage. During the drug intervention period, the general situations of rats in each group were observed and their blood pressure was monitored at specific time points every other week before and after administration. After 8 weeks of drug intervention, enzyme-linked immunosorbent assay was employed to measure the levels of angiotensin-Ⅱ (Ang-Ⅱ) and endothelin-1 (ET-1) in the serum of rats in each group. The corresponding kits were used to determine the levels of nitric oxide (NO), malondialdehyde (MDA), glutathione peroxidase (GPX), superoxide dismutase (SOD), and ferrous ion (Fe2+). Hematoxylin-eosin (HE) staining was employed to observe the changes in the intima of the aorta. Fluorescence quantitative polymerase chain reaction (Real-time PCR) was employed to measure the mRNA levels of PI3K, Akt, Nrf2, heme oxygenase-1 (HO-1), and glutathione peroxidase 4 (GPX4) in the aortic tissue. Western blotting was used to determine the protein levels of p-PI3K (Tyr467/199), PI3K, p-Akt (Ser473), Akt, Nrf2, HO-1, and GPX4 in the thoracic aorta. ResultCompared with the normal group, the model group had decreased body mass, increased irritability, severe endothelial damage, elevated blood pressure and serum levels of Ang-Ⅱ, ET1, MDA, and Fe2+ (P<0.01), lowered NO level (P<0.01), and down-regulated mRNA and protein levels of p-PI3K (Tyr467/199), PI3K, p-Akt (Ser473), Akt, Nrf2, HO-1, and GPX4 in the aortic tissue (P<0.01). Compared with the model group, drug intervention caused no significant change in the body mass, calmed the rats, alleviated the endothelial damage, lowered blood pressure and serum levels of Ang-Ⅱ, ET1, MDA, and Fe2+ (P<0.01), elevated the NO level (P<0.05), and up-regulated the mRNA and protein levels of p-PI3K (Tyr467/199), PI3K, p-Akt (Ser473), Akt, Nrf2, HO-1, and GPX4 (P<0.05). ConclusionThe earthworm protein can exert antihypertensive effects by ameliorating VED in SHR. Specifically, it may regulate the PI3K/Akt/Nrf2 signaling pathway to inhibit oxidative stress and ferroptosis.