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Objective:To explore the effect of repetitive transcranial magnetic stimulation (rTMS) combined with cognitive behavioral therapies (CBT) on the cognitive function and alcohol craving in patients with alcohol dependence.Methods:From March 2019 to September 2021, a total of 150 patients with alcohol dependence were enrolled and randomly divided into rTMS treatment group (rTMS+ sham CBT, n=41), CBT treatment group (CBT+ sham rTMS, n=34), rTMS+ CBT treatment group( n=36) and control group (sham rTMS+ sham CBT, n=39). At baseline (before treatment), 2nd week, 8th week, 12th week and 24th week, alcohol dependence scale (ADS) was used to evaluate the degree of alcohol dependence, the obsessive compulsive drinking scale (OCDS) was used to assess patients' drinking craving, and Montreal cognitive assessment scale (MoCA) was used to assess the overall cognitive level of patients.SPSS 23.0 statistical software was used to compare the differences of ADS, OCDS and MoCA scale scores of the four groups by repeated measure ANOVA and simple effect analysis. Results:(1)The patients in the four groups were evaluated with ADS scale at baseline, 12th week and 24th week respectively.The interaction of group×time( F=1.279, P=0.279) and the main effect of group were not significant ( F=0.882, P=0.454), and the main effect of time was significant ( F=12.925, P<0.001) .Further simple effect analysis showed that the ADS score of rTMS+ CBT group was lower than that of baseline(14.48±5.70, 10.00±6.51) ( P=0.01) at 24th week.(2)Patients in the four groups were assessed with OCDs scale at baseline, 2nd week, 8th week, 12th week and 24th week, and the interaction of group×time was significant ( F=2.015, P=0.042). Further simple effect analysis showed that the OCDs scores of rTMS group and rTMS+ CBT group at each follow-up time node were lower than those at baseline period (all P<0.05). (3)Patients in the four groups were assessed with MoCA scale at baseline, 8th week, 12th week and 24th week, and the interaction of group×time was not significant ( F=1.660, P=0.106), and the main effect of group and the main effect of time were significant ( F=2.964, P=0.038; F=14.239, P<0.001). Further simple effect analysis showed that the score of MoCA scale in CBT group at the 24th week was higher than that at baseline (21.73±5.81, 24.60±3.98)( P=0.029), the score of MoCA scale in rTMS+ CBT group at the 24th week was higher than that at the 8th week (23.50±6.01, 25.95±2.87) ( P=0.006), and the score of MoCA scale in rTMS group at the 12th week was higher than that in control group (22.08±6.64, 26.64±2.46)( P=0.009). Conclusion:rTMS combined with CBT can be effective in improving alcohol craving and cognitive function in patients with alcohol dependence, and has a good long-term effect.
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After traumatic brain injury(TBI), as an immune signal complex in cytoplasm, the assembly and activation of inflammasomes can induce pyroptosis, trigger extensive inflammations, aggravate brain tissue damage, lead to delayed cell death and progressive neurodegeneration, and cause neurological dysfunction, which plays a key role in the development and prognosis of the disease. After TBI, the mechanism of pyroptosis mediated by inflammasomes and its targeted treatments have become the focus of recent researches. The authors review the research progress in inflammasomes-mediated pyroptosis in terms of related signal pathways, their activation and regulation and application of targeted drugs after TBI, so as to provide references for clinical treatments.
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A male patient of acute onset is reported, whose main clinical manifestations were ataxia and dysarthria, with elevated carcinoembryonic antigen, non-small cell lung cancer antigen, carbohydrate antigen 72-4, positive anti-Yo antibody. The patient′s gastroscopy and biopsy result suggested gastric cancer, and his symptoms got better after radical surgery. Anti-Yo-associated paraneoplastic cerebellar degeneration complicated with gastric adenocarcinoma was diagnosed. If encountering cases of ataxia or dysarthria in clinical work, the possibility of paraneoplastic cerebellar degeneration should be considered and evidence for tumor should be searched, so as to avoid missed diagnosis or misdiagnosis.
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Objective: To investigate the ethical decision-making process of breaking confidentiality when counselors dealing with self-inflicted injury and suicide issues in college situation. Methods: A semi-structural interview was addressed to 10 counselors from 7 college counseling centers in Beijing, among whom with (10 ± 8) years of experience on average in this field. Content analysis method was used to transcription of the interviewing data. Results: Totally 8 counselors had received ethical training more or less, and attached great importance to ethical codes. There were still some conflicts between school regulations and confidentiality rules in 7 university counseling centers. Different counselors varied greatly in decision-making on breaking confidentiality when facing college students' self-inflicted injury and suicide. Faced with conflicts between college demands and confidentiality principles, counselors could take the professional standpoint and consider more of the interests of students. Conclusion: The decision-making process on self-inflicted injury and suicide confidentiality breakthrough needs to be standardized. College's attention and support to the counseling work should be strengthen and enhance ethical awareness.
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BACKGROUND:Currently, what we know about exosomes is that it can be produced by a variety of cel s and transfer a variety of materials, producing subsequent function of regulation. OBJECTIVE:To review the function of exosomes in mesenchymal stem cel differentiation, in order to provide reference for further in-depth study. METHODS:With the key words of“exosome, mesenchymal stem cel , differentiation”in Chinese and English, respectively, a computer-based search was performed for articles published in CNKI, Medline and Embase databases from January 2001 to September 2016. After the initial screening, the reserved articles were further detailed, summarized and concluded. RESULTS AND CONCLUSION:Exosomes are a kind of vesicles 40-100 nm in diameter, which can be secreted by a variety of cel types, containing functional products, such as functional protein, gene product, lipid and so on. As a bridge of adjacent cel s transferring functional products, exosomes are becoming an issue of concern in the microenvironment for cel interaction. In the differentiation of mesenchymal stem cel s, exosomes also play an indispensable role via pathway regulation.
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BACKGROUND:The microRNAs are involved in regulation of stem cel proliferation, differentiation and aging. To study the effect of Let-7c, a member of Let-7, on the neural differentiation of bone marrow mesenchymal stem cels provides new ideas for stem cel therapy. OBJECTIVE: To investigate the role of Let-7c in the neural differentiation of bone marrow mesenchymal stem cels. METHODS: The lentiviral vectors of Let-7c-up and Let-7c-inhibition were constructed and transfected into rat bone marrow mesenchymal stem cels. Optimal multiplicity of infection was screened. The cels were divided into non-transfected group, negative control group (transfected with empty virus), transfected enhancement group (transfected with LV-rno-Let-7c-up), transfected inhibition group (transfected with LV-rno-Let-7c-5p-inhibition). Bone marrow mesenchymal stem cels were treated with fasudil as an inducer for triggering the cels to differentiate into neurons. The fluorescence expressed by transfected cels was observed under inverted fluorescence microscope. The expression of neuron-specific markers, neuron-specific enolase and microtubule-associated protein 2, were measured by immunocytochemical method. The mRNA expression of microtubule-associated protein 2 was detected by RT-PCR. The cel viability was determined by MTT method. RESULTS AND CONCLUSION:Under the inverted fluorescence microscope, the cels were successfuly transfected with LV-rno-Let-7c-up and LV-rno-Let-7c-5p-inhibition. Fasudil induced bone marrow mesenchymal stem cels to differentiate into neurons. The transfection efficiency and expression levels of neuron-specific enolase and microtubule-associated protein 2 in the transfected enhancement group were significantly higher than those in the negative control group (P < 0.05), while in the transfected inhibition group, they were lower than those in the negative control group (P < 0.05). These findings indicate that the differentiation percentage of bone marrow mesenchymal stem cels is increased by fasudil after transfection with LV-rno-Let-7c-up, and Let-7c may promote the differentiation of bone marrow mesenchymal stem cels into neurons.
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Bone marrow mesenchymal stem cells (BMSCs),which can be isolated from organs and tissues,are cell populations with high degree of plasticity,similar to hematopoietic stem cells in bone marrow,and can be in vitro cultured,induced and amplified.BMSCs are increasingly used in gene therapy,cell therapy and tissue engineering.BMSCs have been currently studied in a number of autologous transplantations,while not all BMSCs are suitable for autologous transplantation.BMSCs exhibit biological aging gradually when cultured in vitro.Biological aging can be divided into age-induced senescence and long-term passage induced senescence.Aging BMSCs demonstrate changes in biological behaviour which reduces the success rate of autologous transplantation.In this review,biological aging BMSCs are discussed in order to provide help for the transplantation of BMSCs.
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BACKGROUND:There is no clear understanding about the effect of let-7f and interleukin-6 (IL-6) on the proliferation of bone marrow mesenchymal stem cels and their relationship. OBJECTIVE: To explore the effects of expression levels of let-7f and IL-6 on the proliferation of bone marrow mesenchymal stem cels and their relationship. METHODS:(1) LV-rno-let-7f-up and LV-rno-let-7f-down were constructed and transfected into bone marrow mesenchymal stem cels of Sprague-Dawley rats, respectively. Then, there were four groups in the study: transfection upregulation group transfected with LV-rno-let-7f-up), transfection inhibition group (transfected with LV-rno-let-7f-down), negative control group (transfected with FU-RNAi-NC-LV), and untransfected group. The expression level of let-7f in each group was detected by qRT-PCR. The proliferation ability of cels and expression levels of IL-6 when let-7f expression was at different levels were detected by MTT, flow cytometry and ELISA. The expression of Cyclin D1 at mRNA and protein levels was detected by qRT-PCR and western blot, respectively. (2) To predict the potential target gene of let-7f, the wild-type/mutant IL-6 3’UTR reporter gene vectors were constructed, and cotransfected with let-7f/let-7f inhibitor respectively into the 293T cels to measure the luciferase. RESULTS AND CONCLUSION: Compared with the negative control group, the proliferative and cloning capacities of cels in the transfection upregulation group were higher; the number of cels was significantly decreased at G1 stage and increased at S stage, and the apoptotic cels were reduced in number (P 0.05). Luciferase activity of cels transfected with wide-type IL-6 3’UTR and let-7f was significantly reduced (P < 0.05). These findings indicate that up-regulation of let-7f can promote the proliferative and cloning capacities of bone marrow mesenchymal stem cels and reduce cel apoptosis, but downrelation of let-7f exhibits an inhibitory effect. Overexpression of IL-6 can suppress the proliferation of bone marrow mesenchymal stem cels, which is considered to be a target gene of let-7f, and let-7f may suppress the expression of IL-6 to promote the cel proliferation.
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BACKGROUND:MicroRNA plays an important role in the process of growth and aging of living body. To know the role of let-7d in inducing bone marrow mesenchymal stem celldifferentiation into neurons can promote the stem celltransplantation. OBJECTIVE:To investigate the role of let-7d in inducing bone marrow mesenchymal stem celldifferentiation into neurons. METHODS:(1) The lentiviral vector of let-7d was constructed and transfected into rat bone marrow mesenchymal stem cells. The cells were divided into non-transfected group, negative control group (transfected with FU-RNAi-NC-LV), transfected enhancement group (transfected with let-7d-LV), transfected inhibition group ( transfected with let-7d-inhibition-LV). (2) Rat bone marrow mesenchymal stem cells were treated with fasudil as an inducer for triggering the cells to differentiate into neurons. The expression of neuron-specific markers, neuron-specific enolase and microtubule-associated protein 2, were measured by immunocytochemical method. The mRNA expression of microtubule-associated protein 2 was detected by RT-PCR. The viability of bone marrow mesenchymal stem cells was determined by MTT method. RESULTS AND CONCLUSION:Under inverted fluorescence microscope, the cells were successful y transfected with let-7d. Fasudil induced bone marrow mesenchymal stem cells to differentiate into neurons. The transfection efficiency and expression levels of neuron-specific enolase and microtubule-associated protein 2 in transfected enhancement group were higher than those in the negative control group (P<0.05);while in the inhibition group, they were lower than those in the negative control group (P<0.05). These findings indicate that let-7d can promote the differentiation of bone marrow mesenchymal stem cells into neurons induced by fasudil, and by control ing the expression of let-7d we can influence the differentiation efficiency from bone marrow mesenchymal stem cells to neurons.
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Alzheimer disease (AD) is a fatal neurodegenerative disorder,and the most common cause of dementia in the elderly.Accumulated evidences in AD research suggest that alterations in the microRNA (miRNA) network could contribute to risk for the disease.As the second discovered miRNA,Let-7 has an important effect on cell proliferation,differentiation,apoptosis,immune response,tumorigenesis,metastasis,and so on.We review in this article the role of Let-7 family in the pathogenesis of AD to provide a strong basis for future research aimed at understanding the potential contribution of miRNAs to AD pathophysiology.
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Objective To investigate changes in the expression of prepro-orexin and orexin receptor-1 ( OX1R) following permanent middle cerebral artery occlusion ( MCAO ) with or without preconditioning through electrical stimulation of the cerebellar fastigial nucleus (FNS). Methods Wistar rats were subjected to permanent MCAO and randomly divided into 5 groups: a sham-operated control group (PO), an FNS preconditioning + shamoperated control group (FNS-PO) , an ischemia group, an FNS preconditioning + ischemia group (FNS-PI) and a cerebellar fastigial nucleus injury + FNS preconditioning + ischemia group (FNL-FNS-PI). Each group was divided into 5 subgroups according to the time at which the animals were sacrificed after the MCAO ( 1, 3, 6, 12 and 24 h).RT-PCR was used to detect expression of OX1R mRNA, and ELISA to measure the levels of orexin-A in the hypothalamus and plasma. Results The immunoreactivity of prepro-orexin decreased significantly in the PI groups, with further decreases over time. At the 12th h after MCAO, the immunoreactivity of prepro-orexin reached a minimum.There were significant differences between the rats in the PO and FNS-PO groups. On the contrary, the immunoreactivity of OX1R increased significantly in the PI groups, with further increases continuing over time, peaking at 12 h after the MCAO. There were significant differences between the PO and FNS-PO groups. In the rats with FNS preconditioning (PI-FNS) , the decrease in prepro-orexin and the increase in OX1R were significantly inhibited compared to the PI subgroups at the 6th and 12th hour. There was no significant difference between the FNL-PIFNS group and the PI group. The expression of OX1R mRNA increased significantly in the PI group, with further increases continuing over time, peaking at 24 hours. The plasma levels of orexin-A were not significantly different among the groups, but the levels of orexin-A in the hypothalamus decreased significantly in the PI and FNL-PI-FNS groups, with further decreases continuing over time. At the 12th h after the MCAO the levels were significantly different compared with the PO and PO-FNS groups. While in the rats with FNS preconditioning (PI-FNS) , the decrease in orexin-A level was reversed and there was no significant difference compared with PO and PO-FNS groups. Conclusions The orexinergic system is altered following cerebral ischaemia. FNS preconditioning may be able to regulate these changes.
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Objective:To investigate the effects of hyperglycemia on the pancreas and duodenal homeobox gene-1 (Pdx-1) and insulin gene in fetal rat islets in vitro.Methods:Fetal rat islets were digested by collagenase and underwent culture at 5.5 mmol/L, 11.1 mmol/L and 33.3 mmol/L glucose concentration respectively.The insulin content of fetal rat islets and insulin releasing test were measured by radioimmunoassay.The expression of Pdx-1 protein and mRNA were evaluated by indirect immunofluorescence cytochemistry staining and reverse transcriptase-polymerase chain reaction (RT-PCR).Results:The results showed that the insulin content and stimulated index of hyperglycemia group 1 ( 11.1 mmol/L) were significantly higher than that of controls ( 5.5 mmol/L) and hyperglycemia group 2 ( 33.3 mmol/L).The majority of Pdx-1 positive cells in control group were confined to the nuclear periphery, but the majority of Pdx-1 positive cells in hyperglycemia groups were translocated to the nucleoplasm, and the expression of Pdx-1 mRNA was also elevated in hyperglycemia groups.However,the expression of insulin mRNA decresed in hyperglycemia group 2.Conclusion:Hyperglycemia may regulate Pdx-1 in fetal rat islets by increasing its protein level and translocation to the nucleus.
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Objective To identify the effect of decreasing the rejection using the biological semipermeable membrane combined with the privileged site xenotransplantation. Methods The rat ICCs encapsulated by biological semipermeable membrane were xenotransplanted into dog's brain. The pathological changes of implants and surrounding cerebral tissues were observed under light and electric microscopy. The ?-cells of implants were identified by immunohistochemistry. Results After 2 months of transplantation, The ?-cells were observed and the lymphocytes were dispersed in the grafts.The brain tissues near the grafts showed slight edema and glial hyperplasia. Conclusion The method using the biological semipermeable membrane in combination with the privileged site xenotransplantation have a beneficial effect on the inhibition of the rejection of heterogeneous ICCs implant.
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AIM: To investigate whether grafting neural stem cells (NSCs) improves the impaired cognitive deficits and spatial recognition after ischemic-hypoxic brain damage (HIBD) in neonatal rats. METHODS: Non-immunosuppressed 7-day-old SD rats were used as research subject and randomly divided into 3 groups: (1) sham group (n=10); (2) HIBD group (n=11); (3) transplant group (n=13). (2) and (3) were anesthetized and subjected to a hypoxic/ischemic injury obtained by combination of left carotid ligation and exposure to 8% oxygen for 2 h. At 3 days post injury, hypoxic-ischemic brain damaged animals were re-anesthetized and randomized to receive stereotactic injection of NSCs prelabeling with BrdU or control media into the hippocampus in the ipsilateral hemisphere. Cognitive (i.e., learning) deficits were assessed at 2 to 4 weeks after transplantation. At the end of the behavioral tests, the animals were killed and evaluated for NSC survival and histopathological analysis. RESULTS: Transplant group showed significantly improved cognitive function in selected tests as compared with HIBD group during the 4-week observation period. They took less time than HIBD group in finding the 3 arms baited with water and had a decreased number of working and reference memory errors in radial maze acquisition tests. Histological analysis showed that transplanted NSCs attenuated CA1 cell loss after HIBD, and NSCs survived for as long as 4 weeks after transplantation and were detected in the hippocampus. CONCLUSION: These data suggest that transplanted NSCs attenuate brain damage and cognitive dysfunction after hypoxic-ischemic brain damage. This approach warrants continued investigation in light of potential therapeutic uses.
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AIM:To construct a eukaryotic expression vector containing pancreatic duodenal homebox-1(PDX-1)and to elevate the expression efficiency of exogenous gene in rat bone marrow mesenchymal stem cells(MSCs).METHODS:Recombinant vector containing PDX-1 was constructed.Flow cytometry was used to identify the cell cycle of bone marrow mesenchymal stem cells(MSCs)cultured in vitro.Recombinant vector containing PDX-1 was transfected into bone marrow MSCs using superfect in medium.After being selected by G418,RT-PCR and Western blotting were used to investigate the expression of PDX-1 in MSCs.RESULTS:Restricted enzyme analysis and sequencing showed that PDX-1 gene segment was consistent with that in GenBank.Flow cytometry showed that there were about 85.9% cells at the cell cycle of G_0/G_1.The whole cells transfected emitted green fluorescence under flow cytometry.The efficiency of transfection was above 40%.RT-PCR and Western blotting demonstrated that there was expression of PDX-1 in transfected bone marrow MSCs.CONCLUSION:Recombinant vector containing PDX-1 was constructed successfully.Superfect mediated expression of exogenous gene in bone marrow MSCs in a high efficiency,and bone marrow MSCs containing exogenous gene are an ideal cells for gene therapy.
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AIM: To study the protocol and condition that induce bone marrow stromal cells (MSCs) to differentiate into neuron in vitro by baicalin, a kind of flavonoid isolated from an important medicinal plant Scutellariae Radix . METHODS: MSCs from adult rats were induced by baicalin in serum-free medium for 6 h, and postinduced for 6 d. The morphological changes of MSCs were evaluated by light microscope. The positive percentages of neuron-specific enolase (NSE), 200-kilodalton neurofilament (NF), glial fibrillary acidic protein (GFAP) and vimentin expression were measured by immunocytochemistry with ABC staining. Hoechst 33258 staining was used to measure the cell viability by fluorescence microscope. RESULTS: After induction for 6 d, MSCs displayed neuronal morphologies, such as pyramidal cell bodies and processes formed extensive networks. The positive percentages of NSE, NF, GFAP and vimentin protein expression were 70.5%?11.6%, 68.3%?13.4% ,
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Islet transplantation is an effective method for diabetic therapy. This article reviews the recent achievments in the study of induction differentiation of embryonic and adult stem cells into pancreatic islets, and expects that the breakthrough in this field would provide a new method for diabetic therapy.
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Objective To investigate a new method of a long term culture for rat islets in vitro. Methods Rat islets marked by green fluorescent protein (GFP) were cocultured with Sertoli cell for 20 weeks. Histological studies were performed on islet group and coculture group in 1w, 3w, 10w, 15w, 20w of culture by light, fluorescent and electron microscopy. Insulin released was measured by radioimmunoassay. Results In islet group, islet viability and the number of insulin positive cells were significantly decreased after 3w of culture, cumulative quantities of insulin for 24 hours and the stimulation index also fell rapidly under this condition, meanwhile the ultrastructure of islets was destroyed. However, under coculture condition, culture time of islets was prolonged in vitro, islet viability and the number of insulin positive cells were significantly increased, cumulative quantities of insulin for 24 hours and the stimulation index maintained at high level, and the ultrastructure of islets remained normal even after 20w of culture. Conclusion Coculture of rat islets with Sertoli cells may promote islet growth and prolong culture time, and it is a new method of a long term culture of islets in vitro.