1.Effects of hyperbaric oxygen treatment on expression of ERK1/2 and neurological function in rats with traumatic brain injury
Jie SUN ; Bin LING ; Bingqin LI ; Yunfan GU ; Guanghui XIU ; Qiqin DAN
Chinese Journal of Trauma 2011;27(12):1136-1139
Objective To explore the effects of hyperbaric oxygen (HBO) treatment on expression of extra-cellular signal regulated kinase 1/2 (ERK 1/2) and neurological function in acute traumatic brain injury (TBI) rats so as to offer trial support for clinical application of HBO in TBI.Methods Twenty-four adult Sprague-Dawley rats were randomly divided into sham-operation group,TBI group and HBO treatment group,with eight rats per group.The sham-operation group was free from TBI but skull was opened only but the TBI group was subjected to TBI according to Alice method.TheHBO treatment group was treated with HBO for 10 days ( one time per day) after TBI operation.On day 14,NSS rating was performed in all rats.Then,the rats were sacrificed and cortex of which was obtained to measure the expression of ERK1/2 by using reverse transcription-polymerase chain reaction (RT-PCR).The localization of the expression of ERK was detected by immunohistochemical staining and the changes in ERK protein was analyzed by optical density.Results The NSS score in the HBO group was significantly decreased at day 14 after HBO treatment ( P < 0.01 ) and the expression of ERK1/2 in HBO group was also significantly decreased (P < 0.05 ) in comparison with the TBI group.Conclusions HBO treatment can significantly decrease NSS score and expression of ERK1/2 in acute TBI rats,indicating that HBO may improve neurological function through down-regulating expression of ERK1/2.
2.Re-evaluation of indications for prenatal diagnosis of advanced pregnant women under the new screening regime.
Yunfan YANG ; Yanlin WANG ; Jiaqi GAO ; Yi WU ; Jinlin SUN ; Xinrong ZHAO ; Li GAO
Chinese Journal of Medical Genetics 2020;37(10):1057-1060
OBJECTIVE:
To assess the value of non-invasive prenatal testing (NIPT) for women with advanced gestational age but normal measurement for nuchal translucency (NT).
METHODS:
A total of 9371 singleton pregnancies with negative NT screening at early pregnancy were reviewed. Among these, 8627 cases were selected to be screened again by NIPT, and their indications and results were analyzed. The results were compared with those of with other high risk factors and young gestational age.
RESULTS:
The incidence of fetal aneuploidies increased in women with advanced gestational age and ultrasound soft markers, in particular among those who were negative for NT screening but over the age of 37. The detection rate of pathological or likely pathological copy number variations was 1.88% among women who directly underwent invasive prenatal diagnosis because of the advanced age, but there was no correlation with the increase of age. 0.68% of the women where with negative NT screening and NIPT still need to undergo invasive prenatal diagnosis.
CONCLUSION
After NT screening in early pregnancy, NIPT can replace invasive prenatal diagnosis for those below the age of 37, though there is still a possibility of missed detection of pathogenic copy number variation. It is necessary to strengthen ultrasonic monitoring in later period.
3.The expression differences and prognostic value of four cancer stem cell markers in hepatocellular carcinoma tissues and circulating tumor cells in peripheral blood
Hui WU ; Minna SHEN ; Yunfan SUN ; Chunyan ZHANG ; Xinrong YANG ; Beili WANG ; Wei GUO ; Baishen PAN
Chinese Journal of Laboratory Medicine 2021;44(11):1043-1050
Objective:To investigate the expression of four cancer stem cell (CSC) markers (EpCAM, CD133, CD90 and CD24) in hepatocellular carcinoma tissues and peripheral blood circulating tumor cells (CTC),their value in the prognosis of patients with hepatocellular carcinoma.Methods:A total of 50 hepatocellular carcinoma tissues and 29 peripheral blood sample from 50 patients with hepatocellular cancer treated in Zhongshan Hospital Fudan University from October 2013 to September 2014 were collected and analyzed by flow cytometry or qRT-PCR to examine the expression of EpCAM, CD133, CD90 and CD24. The clinical data of patients were collected, including tumor size, tumor number, satellite lesions, vascular invasion, Edmondson stage, BCLC stage and liver cirrhosis, etc. The correlation between the expression of four markers in hepatocellular carcinoma tissues and CTC with the clinical data and survival time of patients were compared.Results:The positive expression rates of EpCAM, CD133, CD90 and CD24 in hepatocellular carcinoma tissues were 66% (33/50), 18% (9/50), 60% (30/50) and 56% (28/50); the positive expression rates in CTC were 55% (16/29), 38% (11/29), 31% (9/29) and 59% (17/29). CD90 expression in hepatocellular carcinoma tissue was positively correlated with the occurrence HCC liver cirrhosis ( P<0.05), while CD133 expression was negatively correlated with the 5-year survival rate of patients ( P<0.05). The expression of EpCAM and CD24 in peripheral blood CTC were closely related to the patient′s Edmondson stage ( P<0.05). The survival time of patients with CD133 positive expression in hepatocellular carcinoma tissue was lower than those without CD133 expression ( P<0.05); the survival rate of patients with EpCAM expressed in either tissue or peripheral blood CTC was lower than that of patients with EpCAM double negative expression ( P<0.05). The survival rate of patients with CD90 negative in HCC tissue and positive in peripheral blood was lower than that in patients with double negative/double positive in tissue and peripheral blood or patients positive in hepatocellular carcinoma tissue and negative in peripheral blood ( P<0.01). Conclusion:Different expression characteristics of four markers in cancer tissues and peripheral blood CTC might provide useful information about predicting prognosis of hepatocellular carcinoma. The expression of CD133 in tissues can be used as an important survival predictor of hepatocellular carcinoma patients. The differential expression of cancer markers in tissue samples and blood samples can provide more clinical prognostic information.
4.Effects of polydopamine-coated porous titanium alloy scaffolds loaded with zoledronic acid-gelatin nanoparticles for topical sustained drug release on osteoclasts: an in vitro study
Xiaojiang YANG ; Guojing SUN ; Weidi YAN ; Zhenfang WU ; Yunfan TI ; Jingwei LU ; Jianning ZHAO
Chinese Journal of Orthopaedic Trauma 2022;24(7):617-623
Objective:To analyze the effects of a novel type of polydopamine (PDA)-coated porous titanium alloy scaffolds loaded with zoledronic acid-gelatin nanoparticles (ZOL-GNPs) for topical sustained drug release on osteoclasts in vitro. Methods:After porous titanium alloy scaffolds were fabricated using electron beam melting technique and ZOL-GNPs with different ZOL concentrations (0, 1, 10, 50, 100, 500 μmol/L) were prepared by desolvation method, PDA-coated porous titanium alloy scaffolds loaded with ZOL-GNPs were constructed by combining the two. The characteristics of the scaffolds were analyzed. The biomechanics of 3 different scaffolds (bare porous titanium alloy scaffolds, PDA-coated porous titanium alloy scaffolds, and PDA-coated porous titanium alloy scaffolds loaded with ZOL-GNPs) were investigated. Drug release detection was carried out by high performance liquid chromatography on the 1st, 4th, 7th, 14th, 21st, and 28th days respectively. The osteoclasts were inoculated into the novel scaffolds with different ZOL concentrations. The expression of osteoclast-related genes was detected by real-time quantitative (RT)-polymerase chain reaction (PCR); the expression of osteoclast-related proteins was detected by Western-blot.Results:The PDA-coated porous titanium alloy scaffolds loaded with ZOL-GNPs were successfully constructed. Electron microscope scanning showed that the GNPs were well spheroidized, smooth in surface, and uniformly dispersed, with a particle size of (243.6±63.4) nm. The ZOL-GNPs were uniformly compounded on the surface and in the pores of the scaffolds, and the spheres were regular in shape with no adhesion. The biomechanical experiments showed that the elastic moduli of the porous titanium alloy scaffolds under 3 different conditions were (1.81±0.12) GPa, (1.80±0.23) GPa and (1.81±0.15) GPa, showing no significant difference ( P> 0.05). The drug release percentage in the porous titanium alloy scaffolds was obviously high on the first day, and increased gradually and slowly in the subsequent 27 days. In the scaffolds with a low concentration ZOL, more osteoclasts adhered and proliferated; in the 50 μmol/L scaffolds, spheroid cells appeared; the spheroid cells increased and even apoptosis occurred with an increase in the ZOL concentration. RT-PCR showed that the expression of Ctsk gene and TRAP gene increased with the increased ZOL concentration, peaked in the 50 μmol/L scaffolds, and then decreased with the increased concentration, showing statistically significant differences ( P < 0.05). Western-blot showed that the expression pattern of Ctsk and TRAP was similar to that of their related genes. Conclusions:The novel PDA-coated porous titanium alloy scaffolds loaded with ZOL-GNPs demonstrate good mechanical properties and an anti-osteoporosis effect via their topical sustained drug release. The scaffolds with a ZOL concentration of 50 μmol/L may exert the best effect on inhibition of osteoclasts.