The aim of this research is to investigate the influence of microencapsulation on the expression of the oxidative stress genes and exogenous regulation of HepG2 cells. We compared the expression of hemeoxygenase-1 (HO-1) and glutathione S-transferases-A1 (GST-A1) in HepG2 cells under different culture conditions through real-time PCR. The effects of exogenous antioxidants on cell viability and albumin levels were also evaluated through MTT assay and ELISA assay. The results showed that after culturing for 6 and 16 days, the expression levels of HO-1 in encapsulated cells were approximately 4.9 and 3.1 times higher than that of monolayer cells at the same culture period; As for the expression levels of GST-A1, they were elevated to 11.2 and 33 times of monolayer cells (P < 0.05). Accordingly, we found that NAC at 5-10 mmol/L significantly increased the viability by 40%-70% and the biosynthetic function by 20%-30% in microencapsulated HepG2 cells (P < 0.05). GSH increased the viability of the encapsulated cells by 20%-55% and the biosynthetic function by 15% (P < 0.05). In conclusion, oxidative stress exists in the microcapsules and affects genes expression. Exogenous antioxidants can prevent the inhibition effects of oxidative stress on cellular growth.
Antioxidants ; pharmacology ; Cell Survival ; Gene Expression Regulation ; Glutathione Transferase ; metabolism ; Heme Oxygenase-1 ; metabolism ; Hep G2 Cells ; Humans ; Oxidative Stress

BACKGROUND: Parkinson disease(PD) is a series of clinical symptom induced by decreased dopamine (DA) in the striatum due to nigral dopaminergic neuronal degeneration. The intracerebral transplantation of secretory DA can reverse or improve the symptoms to a certain extent, but immunologic rejection is still existed.OBJECTIVE: To probe into cell transplantation with immunoisolation in treatment of in rats without application of immunosuppress and observe its mechanical intensity and the biocompatibility of microcapsule .DESIGN: Randomized controlled experiment was designed.SETTING: Biomedical Material Engineering Group, Dalian Institute of ChemicalPhysics , Chinese Academy of Sciences, and Department of Neurology, Second Hospital of Jilin University.MATERIALS: The experiment was performed in Animal Experimental Center of Second Hospital of Jilin University from August 2003 to February 2004, in which, 40 male Wistar rats were employed. PC12 cell was provided from Shanghai Institute of Cellular Biology of Chinese Academy of Sciences.METHODS: 6-hydroxydopamine solution was infused in the striatum to prepare animal model of Parkinson disease. Twenty-five rats of those had been prepared successfully and were randomized into microencapsulated cell transplantation group (12 rats), in which, 25 μL cell-loading sodium alginate-chitosan-solium alginate(ACA)microencapsul suspension (equal to 2.5×104 cells) was injected stereotaxically on two points of the right (affected side) striatum of animal model; non-microencapsulated cell transplantation group (7 rats), in which, 25 μL PC12 cell suspension (equal to 5×104cells) was injected; and empty microcapsul transplantation group (6 rats),in which, 25 μL empty microcapsules suspension was injected . On the 7th day after transplantation, in every group, apomorphine (APO) prepared with saline solution was injected (0.05 mg/kg) subcutaneously in the neck; afterwards, the revolving behavior was recorded for each rat, once per week,totally for 12 weeks. In the 12th week after operation, the rats were sacrificed with anesthesia. The brain tissue was collected for pathological observation and microcapsule were retrieved to evaluation of biocompatibility and immunoisolation.numbers before and after transplantation of each group.RESULTS:Twenty-five rats entered result analysis and the rest was sule: the retrieved ACA microcapsule was integrative in morphology,munoisolation of microcapsule: microencapsuled PC12 cells were prolifercycles before and after transplantation of each group: the records of lateral revolving of rats in every group before transplantation were not significantly different (P ＞ 0.05). In microencapsuled cell transplantation group, 2weeks later, the average number of revolving was significantly lower than that before the transplantation, or even the revolving stopped; the improved symptoms were maintained till the 12th week after transplantation. In nonmicroencapsulated cell transplantation group, the average revolving number was also significantly lower than that before the transplantation, but that on the 8th and 12th weeks was in tendency of increase, without obvious change compared with that before the transplantation (P ＞ 0.05). The revolving number before and after transplantation in non-microencapsulated transplantation group was similar[(10.5±1.4), (10.5±1.3) cyclos/min, P ＞ 0.05].microcapsule provides immune protection. The grafted encapsulated PC12cells survive for along term in the brain of rats with PD, maintain continuously the normal physiological function and improve the symptoms of PD by synthesizing and releasing DA.

Objective To observe the clinical efficacy and toxicity of intraperitoneal chemotherapy combined with intravenous chemotherapy in treatment of cervical cancer.Methods The experimental group of 40 patients received systemic chemotherapy of docetaxel and cisplatin combined abdominal intraperitoneal high-frequency hyperthermia in the control group,40 patients received docetaxel plus cisplatin chemotherapy,The efficacy and toxicity were observed.Results The complete remission rate of observotion group was 82.5％,significantly higher than 62.5％ in control group.Effective rate in observation group was 92.5％ and also significantly higher than 75.0％ in control gronp.After 6 cycles of chemotherapy,the improvement of the experimental group was 85.0％ higher than 57.5 ％ in control gronp.The differemles between the two groups was statistically significant(P ＜ 0.05); the main side effects during chemotherapy are gastrointestinal reactions,since the cells decreased,hair loss,the difference was not statistically significant(P ＞0.05).Conclusion The chemotherapy of docetaxel and cisplatin intraperitoneal chemotherapy can significantly improve the treatment of cervical cancer,without increasing side effects.

ObjectiveTo investigate the effect of microencapsules cells transfected with nerve growth factor (NGF) gene to the sciatic nerve regeneration following sciatic nerve injury in rats.MethodsMicroencapsules containing cells transfected with NGF gene were prepared using drop generative technique and cells were cultured in vitro. Animal model of sciatic nerve cut and sutured was established with Sprague-Dawely rats, and ninety-six animals were randomly divided into group A (in vivo implantation of microencapsules cells transfected with NGF gene), group B (in vivo implantation of microencapsule), group C (in vivo implantation of cells transfected with NGF gene), and group D (negative control group). The nerve conductive velocity (NCV), nerve action potential (NAP), sciatic nerve function index (SFI) were detected in the 4th, 8th and 12th week postimplantation.ResultsThe microencapsules cells transfected with NGF gene in microencapsules retained reliable cell viability and function. The expanded cells formed cell aggregates, with confocal laser scanning microscopy (CLSM), exhibited green fluorescence material in the cell. The NGF concentration in supernatant were arriving at 269 pg/ml when cultured for 10 days. The results of NCV, NAP and SFI tests in group A were higher than those in the other groups (P<0.05).ConclusionAfter implantation, microencapsules cells transfected with NGF gene may secrete NGF continuously in vivo, and has significant improvement effect on nerve regeneration following sciatic nerve injury.

To study the effect of the osmotic stress in the microenvironment on the growth and metabolism of the encapsulated cells under aerobic condition, Osmo-sensitive yeast Y02724 and high-osmotic resistant yeast Hansel were used as models to explore the growth and metabolism state of the cells cultivated inalginate-chitosan-alginate (ACA) microcapsules. The changes of the yeast cells' specific growth rate, maximum product quantity and the secretion of ethanol and glycerol were analyzed. For Y02724, the yield of ethanol was increased in the ACA microenvironment compared to suspension cultivation. For Hansel, the maximum growth speed of microencapsulated cultivation had no obvious difference compared to the suspension cultivation. Moreover, after encapsulation, the production of glycerol was decreased for both Y02724 and Hansel compared to suspension cultivation. In conclusion, osmotic stress existed in the ACA microcapsules and affected the growth and metabolism of the cells.
Alginates ; metabolism ; Capsules ; metabolism ; Cell Culture Techniques ; methods ; Chitosan ; metabolism ; Osmosis ; physiology ; Osmotic Pressure ; Polylysine ; analogs & derivatives ; metabolism ; Saccharomyces cerevisiae ; growth & development ; metabolism ; Yeasts ; classification ; growth & development ; metabolism

Three-dimensional cell culture is a novel type of in vitro culture method.This system can mimic the microenvironment of cell growth in vivo,where cells exhibit similar state and function to that in the in vivo environment.Currently,three-dimensional culture system has been widely applied in tissue engineering and tumor cell biology fields,etc.Radiotherapy is an important treatment of cancer.Radioresistance of tumor cells is the main cause of treatment failure,tumor recurrence and metastasis.Tumor cells can exhibit the resistant characteristics in the three-dimensional culture system,similar to those of tumor cells in vivo.Therefore,three-dimensional culture system can be adopted to investigate the radioresistance and underlying mechanism of tumor cells and identify the key factors regulating the radioresistance of tumor cells,which plays a pivotal role in enhancing the radioresistance and improving the effect of radiotherapy.This article will review recent research progress in the radioresistance and sensitization of tumor cells under three-dimensional culture system,aiming to provide reference for relevant investigations.

Precision medicine is becoming the goal of translational research on colorectal cancer. Accurate molecular subtyping contributes to better guidance of clinical practice. The current TNM staging system of colorectal cancer is inadequate in terms of guiding clinical practice, such as the underestimation of prognosis of with stage II( and III( colorectal cancer TNM staging, and identification of high-risk and low-risk patients with stage II( colorectal cancer. Researchers from Europe and US have proposed a number of molecular subtypings with clinicopathological phenotypes and molecular phenotypes, which has certain practical significance and is beneficial to the choice of treatment regimen and targeted drugs. But the current results of subtyping research require further validations by clinical large scale multi-center trials. Based on precision medicine, molecular subtyping gradually reveals its clinical significance and is optimized through combining genomics with various clinical phenotypes, indicating its guidance for clinical practice, which is the inevitable course of precision medicine accomplishment. In recent years, there have been many new advances in colorectal cancer liver metastasis treatment. The prognosis of colorectal cancer patients undergoing resection of liver metastasis lesion is similar to those with stage III(. Early recurrence within 6 months after translational treatment and resection occurred in about one third of the patients with initially unresectable liver metastasis, and the overall survival was poor. Thus, an evaluation system should be established in order to avoid the strong therapy and strive for better quality of life in some patients. Individualized treatment for colorectal cancer is emphasized increasingly. Body fluid (peripheral blood and urine) marker detection is a recent research hotspot, including serum protein(polypeptide), plasma miRNA, circulating tumor cells and circulating nucleic acid.
Biomarkers, Tumor ; blood ; urine ; Colorectal Neoplasms ; diagnosis ; pathology ; therapy ; Humans ; Liver Neoplasms ; secondary ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Precision Medicine ; Prognosis ; Quality of Life ; Translational Medical Research

Objective:To evaluate the diagnostic value of serum prostate-specific antigen (PSA) levels and multi-parameter magnetic resonance imaging (mpMRI) in patients with granulomatous prostatitis after intravesical Bacillus Calmette-Guérin (BCG) therapy.Methods:The medical records of eight patients with pathologically proven granulomatous prostatitis in Shandong Provincial Hospital Affiliated to Shandong University from January, 2015 to June, 2020, were enrolled and analyzed in this retrospective study. All 8 patients (ages 47-76, mean 63.6) underwent pelvic mpMRI and serum tPSA levels before TURBT, which showed the results of tPSA, f/t and mpMRI were normal before TURBT (0.45-3.62 ng/ml, 0.20-0.51 and normal signal intensities on T1WI and T2WI, respectively). All patients underwent intravesical BCG therapy after post-TURBT 4-6-weeks’ intravesical gemcitabine therapy as a result of pathologically proven middle and high risk NMIBC via cystoscopy.Results:The results of tPSA levels in all 8 patients were elevated after intravesical BCG therapy after 9-15 months (mean 10.5 months), with 4 patients above 4 (6.77-12.89)ng/ml and 4 patients within the normal ranges(2.02-2.68)ng/ml, and f/t levels decreased to lower than 0.16 (0.09-0.15)in all patients. The mpMRI abnormal signals in all patients were all located in the peripheral zone of prostate. All nodular lesions of prostate mpMRI showed lower signal intensity (SI) on T2WI, higher SI on DWI and lower SI on ADC after BCG therapy. All patients underwent prostate biopsy for abnormal signal on prostate mpMRI. The biopsy pathologic results of all patients were granulomatous prostatitis.Conclusions:When elevated PSA and abnormal signals on prostate mpMRI after intravesical BCG therapy occurred, prostate biopsy may not be required for secondary granulomatous prostatitis patients with non-muscle invasive bladder cancer in combination of clinical history.

China ; Counseling ; Female ; HIV Infections ; diagnosis ; Humans ; Pregnancy

Objective:To predict potential target genes in dexamethasone-induced open-angle glaucoma via bioinformatics technology.Methods:The GEO datasets GSE16643, GSE37474, and GSE124114 were used to analyze the differentially expressed genes by GEO2R.Gene Set Enrichment Analysis (GSEA) was performed on the differentially expressed genes between GSE37474 and GSE124114.Intersection of the three datasets were displayed by Venn diagram.The annotation and enrichment analysis of the intersection genes were performed through Gene Ontology (GO)/Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis and then were compared with normal tissue in GTEx Portal database.The corresponding protein interaction network was obtained by STRING.Finally, the candidate genes were searched for their transcription factors in UCSC and JASPAR.Primary human trabecular cells were divided into dexamethasone group and control group, treated with 2 ml 500 nmol/L dexamethasone and the same amount of ethanol, respectively.The expression of BDKRB1 and TAGLN in trabecular cells was detected by Western blot.Results:Differential genes between GSE37474 and GSE124114 datasets enriched in complement and coagulation cascade by GSEA.There were 89 intersecting genes of the three datasets.These genes mainly regulated the formation of extracellular matrix by GO analysis.The gene with the highest enrichment score and collagen-containing extracellular matrix was found to be associated with fibroblasts in GTEx Portal database.ACTA2, MYL9, TAGLN, and LMOD1 were closely related in STRING protein-protein interaction network.Transcription factor SP1 in UCSC and JASPAR according to related genes, BDKRB1, NID1, MFGE8 and TAGLN.The relative expression levels of BDKRB1 and TAGLN proteins were 1.32±0.14 and 0.44±0.09 in dexamethasone group, respectively, which were significantly higher than 1.00±0.00 and 0.20±0.10 in the control group, respectively ( t=-3.61, 2.89; both at P<0.05). Conclusions:Bioinformatics analysis showed that transcription factor SP1 may play a role in human trabecular meshwork cells to myofibroblasts transition after dexamethasone treatment.