1.miR-486 is a tumor suppressor in glioma stem cells
Chinese Journal of Tissue Engineering Research 2015;(41):6633-6637
BACKGROUND:Previous studies have found that the expression level of miR-486 in glioma stem cels (CD133+) is significantly down-regulated compared with that in glioma non-stem cels (CD133-), but the effect of down-regulation of miR-486 on CD133+ cels remains unclear . OBJECTIVE: To explore the effect of miR-486 on CD133+ cels. METHODS:CD133+ glioma stem cels and CD133- glioma cels were separated from U87 cels by flow cytometer. miR-486 overexpression glioma stem cels were constructed by lipofection transfection. RESULTS AND CONCLUSION:After sorting and purification, the content of the CD133+ fraction was enriched up to 83.5%. The expression level of miR-468 in CD133+ glioma stem cels was obviously down-regulated compared with that in CD133- glioma cels. CD133+ glioma stem cels overexpressing miR-486 were fabricated successfuly. Results from in vitro experiments showed that miR-486 overexpression could dramaticaly decrease the proliferation of glioma stem cels, induce a cel cycle arrest in G1/S phase for CD133+ glioma stem cels and promote cel apoptosis. These findings suggest that miR-486 can be a suppressor of glioma stem cels, which offers a novel potential therapeutic target for glioma stem cels and human glioma.
2.Ventriculoperitoneal shunt-related infections caused by Staphylococcus epidermidis
Chinese Journal of Clinical Infectious Diseases 2015;(4):371-375
Staphylococcus epidermidis has the ability to adhere to the surface of shunt tube and to develop the biofilm .It can embed itself in the biofilm so as to escape from the defense of the host .Due to the presence of biofilms , antimicrobial drug resistance , blood brain barrier of central nervous system , the treatment of shunt-related infection caused by Staphylococcus epidermidis is very difficult.This paper reviews the risk factors , prevalence , pathogenesis , treatment and prevention of ventriculoperitoneal shunt-related infections caused by Staphylococcus epidermidis.
3.Study of relationship between levels of hs-CRP and IL-6 with restenosis after stent angioplasty in cerebral arterial stenosis
Changming WEN ; Xinkai WANG ; Baochao ZHANG
Chongqing Medicine 2014;(27):3567-3569
Objective To investigate the relationship between the dynamic changes of hs-CRP and IL-6 with restenosis after stent angioplasty in cerebral arterial stenosis .Methods 65 patients with cerebral artery stenosis stent angioplasty in Nanyang Mu-nicipal Central Hospital from March 2011 to March 2013 were retrospectively analyzed .The changes of hs-CRP and IL-6 levels be-fore operation and at different postoperative time points were observed and their correlation with vascular restenosis was anlyzed . Results (1)The cerebral artery stenosis degree and serum hs-CRP and IL-6 levels were positively correlated(P<0 .05) .(2)Com-paring the hs-CRP and IL-6 levels at different time points showed that the postoperative hs-CRP and IL-6 levels were significantly increased ,reached the highest at postoperative 12 h and then gradually declined .The hs-CRP and IL-6 levels on postoperative 7 d in the patients with restenosis were significantly higher than those in the patients without restenosis (P<0 .05) .(3)The hs-CRP and IL-6 levels at postoperative 6 months in the patients without restenosis were significantly decreased compared with before opera-tion ,but which in the patients with restenosis had no statistically significant differences (P>0 .05);△hs-CRP and △IL-6 had sta-tistically significant differences between the patients with restenosis and the patients without restenosis (P<0 .05) .Conclusion hs-CRP and IL-6 may play an important role in the process of restenosis after stent angioplasty in cerebral arterial stenosis .Monitoring the dynamic changes of hs-CRP and IL-6 has certain value for evaluating vascular restenosis .
4.Collagen-gelatin scaffolds for the repair of peripheral nerve defects
Tiandan LIU ; Baochao ZHANG ; Mingliang HAO
Chinese Journal of Tissue Engineering Research 2017;21(2):286-290
BACKGROUND:Col agen-gelatin composite scaffolds have been reported to be able to promote the early recovery of peripheral nerve defects. However, this conclusion has not been further confirmed. OBJECTIVE:To investigate the biocompatibility of the col agen-gelatin scaffold and its treatment outcomes in the repair of peripheral nerve defects. METHODS:The col agen-gelatin scaffold was co-cultured with bone marrow mesenchymal stem cel s (BMSCs) of Sprague-Dawley rats for 5 days, and then the cel growth was observed. Twenty Sprague-Dawley rats were enrol ed, modeled into a left 30-mm peroneal nerve defect and randomized into experimental and control groups. The col agen-gelatin scaffold composited with BMSCs was implanted into the experimental group, and autograft nerve implanted into the control group. Morphology of the middle bridge was observed, and electrophysiology was conducted at 16 weeks after implantation. RESULTS AND CONCLUSION:BMSCs grew and adhered wel onto the scaffold. The myelinated nerve fiber density did not significantly differ between groups (P>0.05). The myelinated nerve fiber diameter, myelin sheaththickness and percentage of nerve tissues in the experimental group were significantly lower than those in the control group (P<0.05). There were no significant differences in the conduction velocity, latency of motor nerves and the conduction velocity and amplitude of sensory nerves between groups (P>0.05). The amplitude of motor nerves and the latency of sensory nerves in the experimental group were significantly lower than those in the control group (P<0.05). To conclude, the col agen-gelatin scaffold holds a good cytocompatibity and is ideal for the repair of peripheral nerve defects.
5.Effects of midbrain neural stem cells and bone marrow stromal stem cells on behaviors and brain morphology of rats with Parkinson’s disease
Xinming LIANG ; Guohui FU ; Baochao ZHANG
Chinese Journal of Tissue Engineering Research 2015;(36):5838-5842
BACKGROUND:For treatment of central nervous system diseases, neural stem cel s (NSCs) or bone marrow stromal stem cel s (BMSCs) can be transplanted into the brain, but there are less reports to compare the effects of two kinds of stem cel transplantation. OBJECTIVE:To explore the effect of midbrain NSCs and BMSCs on the behavior and brain morphology of rats with Parkinson’s disease. METHODS:Fifty-eight Sprague-Dawley rats were enrol ed to establish Parkinson’s disease models, and then randomly divided into three groups, which were treated with 5μL midbrain NSCs (n=20), 5μL BMSCs (n=20) and 5μL normal saline (n=18) via two coordinate points of the right striatum at 3 weeks after modeling, respectively. At 5 months after transplantation, the rats underwent intraperitoneal injection of apomorphine to observe behavioral changes, and then, the striatum was taken for immunohistochemistry staining. RESULTS AND CONCLUSION:The number of rotations was reduced significantly in the BMSCs and midbrain NSCs groups at 5 months after transplantation (P<0.05), which was significantly lower than that in the normal saline group (P<0.05). However, there was no significant difference between the BMSCs and NSCs groups (P>0.05). In the BMSCs group, BrdU/Nestin positive cel s were seen in the brain stratium at 1 week after transplantation;BrdU/GFAP and BrdU/NSE positive cel s as wel as TH positive cel s rather than BrdU/TH positive cel s were found in the brain stratium at 1 month after transplantation;after that, the number of BrdU/Nestin positive cel s was reduced gradual y and disappeared ultimately, but there were stil a certain number of BrdU/GFAP and BrdU/NSE positive cel s, especial y the former ones. Meanwhile, the NSCs group also had a similar situation, but no double-labeled cel s were in the normal saline group. These findings indicate that midbrain NSCs and BMSCs transplantation can both improve the behavior of Parkinson’s disease rats, and differentiate into neurons, astrocytes and dopaminergic neurons.
6.Paclitaxel and cisplatin inhibit the proliferation of nasopharyngeal cancer stem cells and promote apoptosis via the Wnt/beta-catenin pathway
Yonggang LIU ; Rongsong YANG ; Hongfang WU ; Baochao ZHANG
Chinese Journal of Tissue Engineering Research 2016;20(41):6118-6124
BACKGROUND:Cancer stem cel s have self-renewal ability and can differentiate into new tumors. Cancer stem cel s are the source of tumor formation and recurrence, and they can make tumors insensitive to radiotherapy and chemotherapy.
OBJECTIVE:To explore the effect of paclitaxel plus cisplatin on the proliferation and apoptosis of nasopharyngeal cancer stem cel s (NPCSCs) and involved signal pathways.
METHODS:NPCSCs were sorted by immunomagneticbeads and were treated with paclitaxel, cisplatin or their combination. The expression of caspase-3, activated caspase-3 and Bcl-2, which are related to apoptosis, was determined by western blot. The expression ofβ-catenin and its downstream proto-oncogene, c-myc, was also determined by western blot. The activity of the Wnt/β-catenin pathway was inhibited by knocking downβ-catenin expression orβ-catenin inhibitor XAV939. Proliferation and apoptosis of NPCSCs were detected by MTT and flow cytometry, respectively.
RESULTS AND CONCLUSION:Either paclitaxel or cisplatin could inhibit proliferation and induce apoptosis of NPCSCs. The expression of apoptosis marker, activated caspase-3, was increased and the expression of the inhibitor of apoptosis, Bcl-2, was declined. Combined use of paclitaxel and cisplatin had synergistic effect when used together. Either paclitaxel or cisplatin could inhibit the expression ofβ-catenin and c-myc, suppressed the proliferation and induced the apoptosis of NPCSCs by inhibiting the activity of Wnt/β-catenin pathway. These results indicate that the combined use of paclitaxel and cisplatin may inhibit the proliferation of NPCSCs and promote apoptosis via the Wnt/β-catenin pathway.
7.Protective effect of Baicalin on experimental autoimmune encephalomyelitis in mice
Yingguo REN ; Baochao ZHANG ; Dongpei JIA ; Ke HU
Chinese Journal of Comparative Medicine 2017;27(3):52-56
Objective To observe the effects of different concentrations of baicalin on the mouse model of experimental autoimmune encephalomyelitis (EAE) and to explore its mechanisms.Methods A mouse model of EAE was established with MOG33-55 peptide and bacillus Calmette-Guerin (BGG) vaccine with complete Freund adjuvant (CFA).At the third day after immunization, high and low doses of baicalin were administered to the mice intragastrically once a day for 20 days.The neurological function of mice was evaluated.TUNEL staining was used to detect apoptosis in the spinal cord tissue.The level of ATP in spinal cord tissue was detected by an ATP determination kit.Furthermore, the protein expressions of Bax, Bcl-2, cleaved cas-3 and cleaved cas-9 were detected by western blot, respectively.Results Baicalin improved the neurological function and delayed the onset time in EAE mice.
8.Efficacy and safety of lamivudine in the patients of hepatitis B virus-associated glomerulonephritis treated by prednisolone with leflunomide
Yan ZHANG ; Jiqiang ZHANG ; Baochao CHANG ; Weidong CHEN ; Jing WANG ; Kuihua XU
The Journal of Practical Medicine 2014;(17):2822-2825
Objective To investigate the efficacy and safety of lamivudine in the patients of hepatitis B virus-associated glomerulonephritis (HBV-GN) treated by prednisolone with leflunomide. Methods 41 HBV-GN patients treated by prednisolone and leflunomide were enrolled in this study. According to the presence of the indications of antiviral treatment, the patients were divided into the prevention group and the treatment group received lamivudine treatment, and another 15 patients with chronic hepatitis B were treated with lamivudine as the control group. The biochemical, virological and serological responses during the process of the treatment and the serum levels of IFN-γ and IL-4 were determined. Results No significant differences of biochemical, virological and serological responses in the prevention and treatment groups compared with the control group at 48 weeks after treatment. Levels of the IFN-γ and IFN-γ / IL-4 decreased significantly in the prevention group and treatment groups compared with the control group (P < 0.05, respectively). No serious adverse reactions occurred in all patients. Conclusion The antiviral treatment of lamivudine lead to good curative effect and security in HBV-GN patients received hormones combined with leflunomide treatments, but leflunomide with prednisolone inhibited the production of IFN-γ and removal of hepatitis B virus.
9.Construction and identification of a recombinant PRRSV expressing ORF2 of porcine circovirus type 2.
Tingjie ZHANG ; Xing LIU ; Tao SUN ; Xuejiao ZHU ; Baochao FAN ; Juan BAI ; Ping JIANG
Chinese Journal of Virology 2015;31(1):65-73
Porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) are very two important pathogens that have coursed huge economic losses in swine production in worldwide. In this study,a vector pCMV-TJM containing the full-length cDNA clone of PRRSV attenuated strain TJM-F92 was firstly constructed by PCR method. Then a gene sequence containing Afl II/Mlu I e restriction enzyme sites and a transcription regulatory sequence for ORF6 (TRS6) was inserted be- tween ORF7 and 3'UTR, yielding a expression vector pCMV-TJM-TRS. Subsequently, a plasmid pCMV-TJM-Cap was constructed by cloning of PCV2 ORF2 gene into the unique sites Afl II /Mlu I of pCMV- TJM-TRS plasmid DNA. Then three recombinant PRRSV, rTJM, rTJM/TRS and rTJM/Cap, were rescued by transfection of pCMV-TJM, pCMV-TJM-TRS and pCMV-TJM-Cap into Marc-145 cells, respectively,and confirmed by the genome sequence, restriction enzyme digestion, Western Blot and IFA. They all had the molecular markers which was different from the parent virus. The growth characteristics of the rescued viruses were similar to that of parent virus. rTJM/Cap could also express efficiently PCV2 Cap protein in Marc-145 cells. At passage 8, it still had PCV2 ORF2 gene which examined by RT-PCR. It indicated that the full-length cDNA clone of PRRSV attenuated strain TJM-F92 and recombinant PRRSV rTJM/Cap expressing PCV2 Cap protein were successfully constructed. It made an important foundation for studying on the pathogenic mechanisms of PRRSV and PRRSV-PCV2 vaccine in the future.
Animals
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Capsid Proteins
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genetics
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immunology
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Cell Line
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Circoviridae Infections
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veterinary
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virology
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Circovirus
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classification
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genetics
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metabolism
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Gene Expression
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Open Reading Frames
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Porcine Reproductive and Respiratory Syndrome
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virology
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Porcine respiratory and reproductive syndrome virus
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genetics
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metabolism
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Recombination, Genetic
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Swine
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Swine Diseases
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virology
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Viral Vaccines
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genetics
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immunology
10.Effect of lovastatin on proliferation and apoptosis of glioma stem cells
Gongling WEN ; Changming WEN ; Yanping WANG ; Meijuan KANG ; Jing ZHOU ; Baochao ZHANG
Chinese Journal of Tissue Engineering Research 2016;20(32):4778-4784
BACKGROUND:Increasing evidence has shown that lovastatin with less toxicity to normal cels has crucial effects on proliferation, apoptosis and differentiation of various cancer cels. However, its roles in glioma stem cels remain unclear.
OBJECTIVE:To explore the effect of lovastatin on proliferation and apoptosis of glioma stem cels.
METHODS: Flow cytometric sorting was used to separate glioma stem cels from human glioblastoma cel line U87. Effects of lovastatin on the proliferation and apoptosis of glioma stem cels were determined by MTT and flow cytometry, respectively. Furthermore, expression levels of Ki67, Bax and Bcl-2 in glioma stem cels treated with lovastatin were detected using western blot analysis.
RESULTS AND CONCLUSION: The CD133-positive glioma stem cels were sorted from human glioblastoma cel line U87 with a positive percentage of 85%. MTT assay showed that lovastatin inhibited the proliferation of glioma stem cels in dose (5, 10, 20 μmol/L)- and time (24, 48, 72, 96 hours)-dependent manners. Flow cytometry analysis showed that 10 μmol/L lovastatin (48 hours) induced apoptosis in glioma stem cels. In addition, the expression level of Ki67 was decreased by lovastatin treatment in a dose-dependent manner, and the Bcl-2 and Bax expression levels were reduced and increased by 10 μmol/L lovastatin treatment, respectively. In conclusion, lovastatin can inhibit cel proliferation and induce apoptosis of glioma stem cels, and lovastatin may be a potential drug for treatment of brain tumors.