2.Hepatocyte growth factor in combination with interleukin-6 to induce the in vitro differentiation of mesenchymal stem cells isolated from human umbilical cord blood into hepatocyte-like cells
Hong YUAN ; Yuzhuo WU ; Hong CHEN
Chinese Journal of Tissue Engineering Research 2007;0(12):-
AIM: Mesenchymal stem cells (MSCs) isolated from human umbilical cord blood can differentiate into hepatocyte-like cells under a suitable induction condition. The optimal condition is still unclear. This study investigated the feasibility of differentiation of MSCs isolated from human umbilical cord blood into hepatocyte-like cells with hepatocyte growth factor (HGF) and interleukin-6 (IL-6) in vitro to find a new cell source for live tissue engineering. METHODS: The experiment was performed from March to September 2007 at the Central Laboratory of First Hospital of Lanzhou University. ①Three cases of umbilical cord blood were collected from full-term pregnant women. Pregnant women had signed an informed consent. The experiment was approved by Hospital’s Ethical Committee. ②MSCs from umbilical cord blood were separated by density gradient centrifugation and adherent method. Cell surface molecule was measured by flow cytometry. Third passage of cells were divided into four groups and cultured in DMEM medium with HGF (HGF group), IL-6 (IL-6 group), HGF+IL-6 (HGF+IL-6 group) or no growth factor (control group). ③The characteristics of proliferation and growth of MSCs were studied by microscopy and methyl thiazolyl tetrazolium (MTT). The phenotypes of MSCs were identified by flow cytometry and immunohistochemical method. Albumin levels in culture supernatants were determined with enzyme-labeled immunosorbent assay (ELISA). RESULTS: ①Growth and division of adherent cells obtained from human umbilical cord blood were good. The shape of MSCs changed into triangle, polygonal or round on days 21-28 in induction groups. ②Positive staining reaction for alpha fetoprotein (AFP) on day 7, for CK18 on days 21, 28 after induction. Albumin production by induced MSCs increased in a time-dependent manner. The positive percents of every hepatocytic marker were higher in the HGF+IL-6 group than in the IL-6 group and HGF group (P
3.Study on the reversion of drug resistance in human cervical cancer cell lines
Hong CHEN ; Huizhen CHEN ; Xufeng WU
Chinese Journal of Obstetrics and Gynecology 2000;0(11):-
Objective To determine the resistance reversion of mitomycin (MMC) by 3′-Keto-bmt 1-val 2-cyclosporin (SDZ PSC 833) in human cervical cancer in vitro and in vivo. Metheds A xenografted mitomycin resistant mice model of cervical cancer was devolped. The reversion of mitomycin resistance by SDZ PSC 833 (1 or 3 mg/L) was detected from human cervical cancer cell (Hela) and its resistant subline Hela/MMC in vitro and in vivo. Studies in vitro include drug resistance reversion experiment and the changes of morphology. Studies in vivo including tumor volume and tumor related histopathological changes in the autopsied specimen were evaluated by comparing random sections of each group. Results Nontoxic doses of SDZ PSC 833 could result in almost partial reversion of MMC-resistance of Hela/MMC.In vivo studies also showed SDZ PSC 833 augmented the growth inhibitory effect of mitomycin on Hela/MMC xenografted in nude mice. Conclusion SDZ PSC 833 can overcome mitomycin resistance of Hela/MMC in vitro and in vivo ,so SDZ PSC 833 will be a better candidate clinically for reversing multidrug resistance.
4.Ring chromosome 20 syndrome.
Chinese Journal of Pediatrics 2013;51(9):710-712
Anticonvulsants
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administration & dosage
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therapeutic use
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Brain
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diagnostic imaging
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physiopathology
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Child
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Child, Preschool
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Chromosome Deletion
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Chromosome Disorders
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diagnosis
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genetics
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Chromosomes, Human, Pair 20
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genetics
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Electroencephalography
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Epilepsy
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diagnosis
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drug therapy
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genetics
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Epilepsy, Complex Partial
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diagnosis
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drug therapy
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genetics
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Humans
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Karyotyping
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Radiography
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Ring Chromosomes
5.Construction of recombinant eukaryotic expression vector pVAX-S_2S containing the gene of hepatitis B virus surface antigen and DNA-mediated immunization
Hong WU ; Guangming CHEN ; Rude YANG
Chinese Journal of Immunology 1999;0(12):-
Objective:In order to obtain safer HBV DNA vaccine,recombinant kanamycin resistance eukaryotic expression plasmid containing the gene of HBV surface antigen was constructed and used to study humoral immune response in BALB/c mice.Methods:HBV antigen middle protein(preS2+S) encoding gene fragment was isolated from the recombinant eukaryotic expression plasmid pcDNA-S 2S;subcloned into eukaryotic expression vector pVAX1.After confirmed the presence of the gene by restriction endonuclease analysis,it was used to immunize the healthy BALB/c mice at different doses,and the levels of anti-HBs in serum was determined by ELISA.Results:Restriction endonuclease analysis confirmed recombinant plasmid pVAX-S 2S was what was expected.Serum anti-HBs was detected at the 2nd week after DNA vaccination at all three doses(100 ?g,50 ?g,10 ?g per mice)and their titers were increased with time.Quantitative comparison of the serum anti-HBs levels revealed significant(P
6.Protective effect of iscbemic preconditioning on pre-injured heart in rats
Xudong WU ; Chengbin XU ; Hong CHEN
Chinese Journal of Interventional Cardiology 1996;0(04):-
On the model of isoprenaline (ISO)-perinjured rat heart, the protctive effect of ischemic preconditioning (IPC) on ischemia reperfusion (I/R) damage was observed.It was found that compared with alone I/R group,IPC ameliorated I/R-induced reduced reduction of coronary blood flow (CBF) (8. 8 + 0. 6 vs 6. 6 + 0. 4mL/min,P
7.Suppression of the expression of exo- or endogenous genes by shRNA with the aid of PCR
Quan HONG ; Di WU ; Xiangmei CHEN
Medical Journal of Chinese People's Liberation Army 2001;0(09):-
Objective To investigate the effects of shRNA produced by PCR on the suppression of the expression of exo- or endogenous genes. Methods The specific primers were designed, with which the shGFP-RNA and shFN-RNA were produced by PCR. The shGFP-RNA was transfected into 293T cell lines together with pEGFP plasmid, then the cells were detected by laser confocal microscopy 48h later. The shFN-RNA was transfected into rat mesenteric cell lines, then the cells were collected 48h later and the total RNA was extracted, which was reversely transcripted to cDNA. Then the expression level of FN mRNA was examined with real-time PCR, and the expression level of FN protein was examined with Western blot analysis. Results The results of laser confocal microscopy indicated that the EGFP could be successfully suppressed by shGFP-RNA produced by PCR; the results of real-time PCR and Western blot analysis revealed that FN expression level of the cells transfected with shFN-RNA was down regulated, and the level was much lower than those tansfected with independent shRNA (P
8.Nursing of cancer patients with Wernicke encephalopathy after gastrointestinal surgery
Ying WU ; Ruyi YANG ; Hong CHEN
Modern Clinical Nursing 2016;15(8):38-41
Objective To summarize the experience of nursing cancer patients with Wernicke encephalopathy (WE) after gastrointestinal surgery. Method The clinical data of 6 patients with WE after gastrointestinal surgery in our hospital from January 2005 to June 2015 were retrospectively analyzed to summarize the nursing strategies. Results All the 6 patients showed mental changes of different types, 5 patients developed with ocular manifestations and one patient with ataxia. Only one patient showed the classic triad of the disease at clinical presentation. All the 6 patients had characteristic manifestations on cranial MRI and received thiamine (vitamin B1) at diagnosis. Four patients were completely recovered and 2 were partially recovered. Conclusion Timely sufficient thiamine supplementation during the perioperative period is the key to prevent and treat WE in the patients with gastrointestinal cancer. Careful observation of the patients′ mental state and the effective nursing for the special classic triad of the disease are critical for a better prognosis.
9.Role and bioinformatics analysis of UCP2 in cervical cancer
Jing JIANG ; Yan CHEN ; Hong WU
The Journal of Practical Medicine 2017;33(1):91-94
Objective To investigate the effect of UCP2 on cell proliferation, apoptosis, metastasis in cervical squamous carcinoma. Methods Plasmid?mediated downregulation of UCP2 was obtained in cervical cancer cell lines. MTT, flow cytometry and transwell chamber assay were conducted to detect the ability of proliferation, apoptosis and metastasis. Characteristics of UCP2 protein was analyzed by bioinformatics methods. Results (1)UCP2 was verified to be downregulated by qRT?PCR and Western blotting assay.(2)MTT, apoptosis assay and transwell chamber assay showed that the proliferation of SiHa cell was significantly inhibited, and the apoptosis rate was significantly increased, and metastasis was markedly deduced (P < 0.05). (3) Bioinformatic analysis showed that UCP2 was located in mitochondria with several phosphorylation sites, and UCP2 interacted with other proteins to produce biological effects. Conclusion UCP2 may play an important role in the occurrence and development of cervical cancer, and it is expected to be a new target for the early diagnosis and treatment of cervical cancer.
10.Optimized preparation of Silymarin Dropping Pill by a central composite design-response surface method
Chinese Traditional and Herbal Drugs 1994;0(05):-
Objective Using solid dispersion technique to prepare Silymarin Dropping Pill to accelerate dissolution and to improve bioavailability. A central composite design-response surface method was employed to select the optimum formulations. Methods Independent variables were Poloxamer 188 content and silymarin content, while dependent variables were disintegrating time and percent of silymarin dissoluted at a definite time. Linear, two and three order quadratic models were used to estimate the relationship between independent and dependent variables. Response surfaces were delineated according to best-fit mathematic models and optimum formulations were selected there from. Prediction was carried out through comparing the observed and predicted values. Results Three order quadratic equation was the best-fitted mathematic models to describe the relationship between dependent and independent variables, with a regression coefficient of 0.998. Bias between observed and predicted values of disintegrating time and dissolution percentage of optimum formulation dropping pill were negligible, indicating the high predictability of the fit models. Percent dissolution of Silymarin Dropping Pill at 60 min was 19 times as that of conventional tablets. Conclusion Dissolution speed of silymarin can be effectively improved through incorporating into dropping pills. It shows that the optimum mathematic model is highly predictive. The central composite design-response surface method can be fairly used in formulation screening.