4.Experimental study of biomimetic nanoparticle probe containing hematoporphyrin monomethyl ether and its imaging and sonodynamic therapy for breast cancer
Xiaohong LIN ; Pan LI ; Qianru LI ; Ju HUANG ; Qinqin JIANG ; Li WAN ; Rui TANG ; Tao HE
Chinese Journal of Ultrasonography 2021;30(2):173-179
Objective:To develop a biomimetic nanoparticle probe of hematoporphyrin monomethyl ether (HMME) coated with breast cancer cell membrane, to observe its ability to target homologous breast cancer cells in vitro, and to investigate its effect of enhanced photoacoustic imaging and sonodynamic therapy (SDT) for breast cancer in vitro.Methods:The cell membrane of breast cancer 4T1 was extracted by chemical cleavage and repeated freezing and thawing. Then the HMME-coated polylactic acid-glycolic acid copolymer biomimetic nanoparticle was prepared by double emulsification and extrusion. The basic characteristics of nanoparticles were detected. The target ability of nanoparticles to homologous breast cancer cells and the enhancement of photoacoustic imaging were observed in vitro. Singlet oxygen sensor green (SOSG) was used to verify the reactive oxygen species (ROS) production of nanoparticles, and its SDT effect on breast cancer cells was evaluated by CCK8 cytotoxicity assay.Results:The size of the prepared CHP-NPs was uniform, the morphology was spherical "core-shell structure" , the particle size was (275.23±8.25)nm, and the surface potential was (-18.43±0.45)mV. It was observed that CHP-NPs could target homologous 4T1 cells under laser confocal microscopy. In vitro photoacoustic imaging experiments show that the photoacoustic signal of nanoparticles increases with the increase of its concentration. According to SOSG probe detection, CHP-NPs could produce ROS under ultrasonic irradiation.When CHP-NPs was incubated with 4T1 cells alone and no ultrasonic irradiation was used, the cell survival rate was not significantly affected. When the concentration was 0.6 mg/ml, the cell survival rate was still 95%. After ultrasonic irradiation, CCK8 experiment showed that the CHP-NPs had a significant SDT effect on breast cancer cells.Conclusions:The biomimetic nanomolecular probe of breast cancer cell membrane is successfully prepared. The probe has good ability to target homologous tumor, and can significantly enhance tumor photoacoustic imaging and SDT effect.
5.Long-term follow-up for neurogenic bladder treated by sacral neuromodulation: 1 case report
Guang FU ; Limin LIAO ; Zongsheng XIONG ; Chunsheng JU ; Yanhe JU ; Dong LI ; Wenbo SHI ; Juan WU ; Yue HUANG
Chinese Journal of Rehabilitation Theory and Practice 2005;11(11):901-902
ObjectiveTo explore the efficacy and safety of sacral neuromodulation(SNM) for the treatment to neurogenic bladder.MethodsOne patient with neurogenic bladder after spinal bifida underwent the therapy of SNM 42 months ago.The therapeutic efficacy was evaluated and followed up by means of the symptom improvement and voiding diaries.ResultsDuring the test stimulation period,there were significant improvements(>50%) in the objective findings and subjective symptoms.This patient received permanent electrode and neurostimulator implantation and lower urinary tract symptoms were improved continuously until 42 months.ConclusionSNM may be effective for some neurogenic dysfunctions of the bladder.
6.The construction of dicistron system to improve the expression level of bFGF in Escherichia coli
Gengsheng HUANG ; Ju WANG ; Fenyong SUN ; Zhiying LI ; Ling ZHANG ; An HONG ; Guishen LI
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To investigate the condition necessary for high-level expression of basic fibroblast growth factor (hbFGF) in Escherichia coli, a dicistron system was constructed. METHODS: The phage T7 gene ?10 N-terminal sequence, present in the expression plasmid pET-3c, was incorporated into the first cistron, the coding region for bFGF was incorporated into the second, and both were under the control of a transcription promotor recognized by the bacteriophage T7 RNA polymerase. RESULTS: Recombinants were transformed into BL21(DE3) and induced by IPTG for expression, accumulation of up to 20% of the total cell protein of bFGF were obtained, showing bioactivity indistinguishable from standard protein. CONCLUSION: It suggested that such dicistron system was effective for enhancing hbFGF expression.
7.Effects of Pseudomonas aeruginosa vaccine on proliferation and invasiveness of hepatocellular carcinoma cell line MHCC97L
Jianwei ZHOU ; Tao LI ; Zhenggang REN ; Binbin LIU ; Xiuyan HUANG ; Minjie JU ; Zhaoyou TANG
Chinese Journal of Hepatobiliary Surgery 2010;16(6):455-459
Objective To investigate the effects of Pseudomonas aeruginosa vaccine (PA) on proliferation and invasiveness of the hepatocellular carcinoma cell line MHCC97L with metastatic potential. Methods Proliferation, growth curve, plate efficiency, flow cytometry, transwell invasion assay, cell motility assay, scarification test, vascular endothelial growth factor (VEGF) and matrix metalloproteinase-2 (MMP2) protein activity were evaluated after cells were treated with PA at various concentrations. Results PA can inhibit the proliferation and plate efficiency of MHCC97L cell markedly in a dose-dependent manner. The IC50 of cells treated with PA for 48 h and 72 h was 3.1 ×108/ml and 1.9 × 108/ml, respectively. The doubling time increased and plate efficiency decreased gradually when cells treated with 0.5 × 108/ml, 1 × 108/ml and 2 × 108/ml PA (P<0.01). PA could induce cell cycle arrest at the G1 phase in a dose-dependent manner by flow cytometric analysis. The average amount of invading cell per field in cell invasion assay and motility assay were 4. 8 ± 1.3 and 8. 8±2.2 when cells treated with 1× 108/ml PA, which was significantly lower than that of control group (8. 6±2. 1 and 15. 6±1.2 ) (P<0.01) Scarification test showed that the metastatic ability of cells treated with 1 × 108/ml PA significantly lower than that in the control group. Comparison between cells treated with 1 × 108/ml PA and control group, no remarkable difference was found regarding expression of VEGF and MMP2 in supernatant of cell culture. Conclusion PA can inhibit proliferation and plate efficiency of HCC cell line MHCC97L, which is in part mediated by the cell cycle arrest at the G1 phase. PA could inhibit invasiveness of HCC cell line MHCC97L, which is unrelated to the VEGF and MMP2 protein activity.
8.Accurate Determination of 13 C Isotopic Abundances of Free Intracellular Amino acids with Low Concentration by GC-MS-Selective Ion Monitoring Method
Minchao LI ; Mingzhi HUANG ; Yuwei LIU ; Ju CHU ; Yingping ZHUANG ; Siliang ZHANG
Chinese Journal of Analytical Chemistry 2014;(10):1408-1413
13 C isotopic abundance of intracellular free amino acid with a characteristic of fast- turnover can quickly reflect changes in intracellular metabolic state. But the concentration of intracellular free amino acid is low, the existed 13 C isotope detection method based on GC-MS can not satisfy the requirement with full scan mode. In this study, the selected ion monitoring method was used to detect accuracy higher likelihood of analysis of 13 C isotopic abundance of free intracellular amino acid. First, in the full scan mode we analyzed of the fracture law of different amino acids, found the feature corresponding to each amino acid fragments, and established 16 kinds of free intracellular amino acids characteristic fragment library. Then using this characteristic fragment library, only specific m/z signal was detected in sample analysis, which realized the selected ion monitoring and improved the quality of signal. The results of amino acid standards showed that the signal-to-noise ratio, measurement precision and accuracy were improved by 17, 2. 0 and 3. 8 times compared with the full scan mode. In the analysis of coenzyme Q10 producing strains of samples, this method was successfully used to detect isotopic abundance of 8 kinds of free intracellular amino acids. This method plays an important role in the detection of 13 C isotopic abundance of the intracellular free amino acid in cell metabolism research.
9.Influence of early rapidly increased amino acid dosaging on nitrogen balance and growth in preterm infants
Enping XIE ; Jianhua SUN ; Yi SHEN ; Huiqun JU ; Jing LI ; Guoqing ZHANG ; Ping HUANG
Chinese Journal of Clinical Nutrition 2014;22(3):136-140
Objective To explore the influence of early rapidly increased amino acid dosaging on the nitrogen balance and growth in preterm infants.Methods A total of 37 prematures admitted to our NICU within 24 hours after birth and with birth weight of 1 000 g to 1 800 g from November 2011 to January 2013 were randomly divided into study group (n =18) and control group (n =19).Parenteral nutrition (PE) was provided in both groups within 24 hours after birth.The starting amino acid dose was 1.5g/kg per day,then rose by 1 g/kg per day in study group and by 0.5 g/kg per day in control group,until to maximum of 3.5 g/kg per day.The dosage of lipids,glucose,and electrolytes in parenteral nutrition was provided routinely.The trial was double-blinded.Results In the first 5 days of PE,the nitrogen balance had no significant difference between these two groups (F =0.025,P =0.652) ; however,the time of regaining birth weight was significantly shorter in study group [(7.23±2.48) d] than in control group [(9.42±2.03) d] (t=-2.090,P=0.044).There were no differences between these two groups in trems of completely feeding time,weight gain rate,head circumference growth rate,length of hospital stay,and hospital costs (all P > 0.05).Meanwhile,the rates of neonatal necrotizing enterocolitis,sepsis,mechanical ventilation administration,and damage of hepatic and renal function were also not significantly different.The administration of different dosages of amino acids did not affect the energy intake in both groups (F =1.893,P =0.218).Conclusions Prematures need early amino acid administration,with a starting dosage of more than 1.5 g/(kg · d).Rapid increase of amino acid dosing will not affect early nitrogen balance in premature infants; meanwhile,it contributes to weight gain without increasing the complications and hospitalization costs.
10.Comparison between CT/MR-contrast enhanced ultrasound image fusion and contrast enhanced ultrasound in ;immediate assessment for hepatocellular carcinoma ablation
Jinxiu JU ; Kai LI ; Rongqin ZHENG ; Erjiao XU ; Zhongzhen SU ; Dongmei HUANG
Chinese Journal of Ultrasonography 2015;(6):508-511
Objective To evaluate the clinical value of CT/MR-contrast enhanced ultrasound (CEUS) image fusion in immediate assessment for hepatocellular carcinoma (HCC)ablation.Methods Patients diagnosed with HCC to receive ablation therapy were randomly divided into two groups.The ablation for each patient was performed according to their pre-ablation plan.Immediate assessments for the ablation were performed using CT/MR-CEUS image fusion (IF group)and CEUS (CEUS group)separately and whether the need for additional ablations was decided and performed during the therapy.The therapy efficacy was evaluated with enhanced CT or MR 1 month after each complete ablation treatment.Results There were 101 patients with 131 nodules in IF group,and 15.1% of the nodules needed additional ablation according the immediate assessment during the therapy.The complete ablation rate of IF group was 99.2%(125/126).On the other hand,there were 93 patients with 122 nodules in CEUS group,and 4.2% of the nodules needed additional ablation according the immediate assessment during the therapy.The complete ablation rate of CEUS group was 94.2% (113/120).IF group has significantly higher additional ablation rate and complete ablation rate than CEUS group (P <0.05).Conclusions CT/MR-CEUS image fusion can further improve ablation efficacy with accurate intraoperative evaluation and guidance of additional ablation compared with CEUS.