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.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.
6.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.
7.Construction and expression of a pEGFP-C2-HDAC2 recombinant plamid
Hui ZHANG ; Cheng HUANG ; Erbao BIAN ; Bin ZHAO ; Baoming WU ; Changwei LIU ; Xiaoxia CHEN ; Ju LI
Chinese Pharmacological Bulletin 2014;(6):812-815,816
Aims HDAC2 gene was cloned into pEGFP-C2 vector to explore the efficiency of the plasmid trans-fection in renal fibroblasts COS-7 cells to identify the expression of both mRNA and protein levels and to ob-serve the distribution of the protein. Methods The HDAC2 cDNA was amlified by PCR and cut with the double enzyme Xho I and BamH I, then inserted into the eukaryotic expression vector pEGFP-C2 with T4 en-zyme. The recombinant vector was verified by PCR, restriction enzymes cut and sequencing identification. Then it was transfected into COS-7 cells and the ex-pression of pEGFP-C2-HDAC2 was monitored by fluo-
rescence microscope and PCR. Results Fragments of HDAC2 could be seen after dealt with double diges-tion, and GFP could also be detected in the transfected COS-7 cells. HDAC2 gene expression could be detec-ted by PCR and Western blot. The fusion expression of pEGFP-C2-HDAC2 could be detected by Western blot. Conclusion Eukaryotic expression vector of HDAC2 has been successfully constructed, the fusion expres-sion of HDAC2 and GFP protein can be detected in COS-7 cells.
8.X-ray diagnostic site selectivity studies of skeletal fluorosis
Heng-xiang, LI ; Xue-song, WANG ; Pei-zhong, CHEN ; Jie, GAO ; Ju-mei, HUANG
Chinese Journal of Endemiology 2013;32(5):565-568
Objective To study the X-ray signs of forearm and leg in skeletal fluorosis and its diagnostic value,aim at finding the easy examination parts.Methods One thousand four hundred and forty subjects were examined using developed shield,darkroom and other portable dedicated device combined with a small X-ray machine.A total of 384 cases were diagnosed skeletal fluorosis.All patients were divided into different groups and the time,degree and range of X-ray to the forearm and calf elbow,knee,and long bone were compared.Results The X-ray change in the forearm elbow was earlier than that of the leg knee,and trabecular bone change was the earliest indicator,197 cases and 157 cases,respectively,and the difference was statistically significant (x2 =28.006,P < 0.01).Membrane ossification of forearm backbone was earlier than that of the leg,and most of them were degree Ⅰ photos,213 cases and 126 cases respectively.The difference was statistically significant (x2 =17.626,P < 0.01).The direction of the interosseous membrane ossification was from the forearm radius to the ulna,then to the fibula and tibia,and was accompanied by changes in the aggravation of forearm.A variety of indicators were observed,especially the membrane ossification in bone and joint trabecular bone and the long bone was the most active,and the forearm was more sensitive,obviously than that of the calf.Conclusion In the X-ray screening or detection of endemic fluorosis,the forearm radiography is a simple,economical,and effective diagnostic method.
9.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.
10.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.