ABSTRACT
Background Fluorine accumulates in the brain tissue after long-term excessive intake and subsequently cause nerve damage and decline of learning and memory ability. Receptor of advanced glycation end-products (RAGE)/p38 mitogen-activated protein kinase (p38MAPK)/nuclear factor kappa-B (NF-κB) signaling pathway is considered to be involved in the associated mechanism. Objective To study the changes of RAGE/ p38MAPK/ NF-κB signaling pathway in rats with subchronic fluorosis, and to explore the protective effects of extract of Ginkgo biloba 761 (EGb761) and RAGE antagonist (FPS-ZM1) on neuromemory ability. Methods Ninety male clean SD rats were divided into 9 groups with 10 rats in each group. The modeling period was 6 months. Control group (C group): free drinking tap water (fluoride content <0.5 mg·L−1), low- and high-dose fluoride groups (LF group, HF group): free drinking tap water with 10 or 50 mg·L−1 fluoride; intervention group of Ginkgo biloba extract (CE, LFE, and HFE groups): on the basis of the C group, LF group, and HF group, 100 mg·kg−1·d−1 EGb761 was given daily via intragastric administration; FPS-ZM1 intervention groups (CF, LFF, and HFF groups): 7 d before the end of modeling, 1 mg·kg−1·d−1 FPS-ZM1 was injected intraperitoneally daily on the basis of the C group, LF group, and HF group. The contents of fluoride in brain and blood of each group were detected. The learning and memory ability was tested by water maze experiment. The histopathologic changes of the hippocampus were detected by Nissl staining. The protein expression levels of RAGE and its ligand high mobility group protein B1 (HMGB1), NF-κB, p38MAPK, phospho-p38MAPK (p-p38MAPK), interleukin-6 (IL-6), and tumour necrosis factor-α (TNF-α) in brain tissue were detected by Western blotting. The mRNA expression levels of RAGE, HMGB1, and p38MAPK were detected by quantitative real-time PCR. Results Compared with the C group, the contents of blood fluoride and brain fluoride in the LF and the HF groups were increased (P<0.05). The results of the water maze experiment showed that, compared with the C group, the escape latency time of the LF group and the HF group was longer and the crossing times were reduced; compared with the HF group, the escape latency time of the HFE group and the HFF group was shortened, and the crossing times were increased (P<0.05). The Nissl staining results showed that the number of Nissl body in the HF group decreased compared with the C group; compared with the HF group, the number of Nissl body in the HFE group and the HFF group increased. The Western blotting results showed that compared with the relative protein expression levels of RAGE, HMGB1, NF-κB, p38MAPK, p-p38MAPK, IL-6, and TNF-α in the C group , the levels of above indicators in the HF group and the levels of RAGE, HMGB1, NF-κB, p-p38MAPK, and IL-6 in the LF group were up-regulated (P<0.05); compared with the HF group, the levels of above indicators in the HFE group and the HFF group were all down-regulated (P<0.05); compared with the relative protein expression levels of RAGE and HMGB1 in the LF group, the levels in the LFE group and the LFF group were all down-regulated (P<0.05). The quantitative real-time PCR results showed that compared with the C group, the mRNA expression levels of RAGE and HMGB1 in the LF group and the HF group were up-regulated; compared with the LF group, the mRNA expression levels of RAGE in the LFE group and the LFF group were down-regulated ; compared with the HF group, the mRNA expression levels of RAGE and HMGB1 in the HFE group and the HFF group were down-regulated (P<0.05). Conclusion The central nervous system injury caused by subchronic fluorosis may be related to the activation of RAGE/p38-MAPK/NF-κB signaling pathway, which can impair the learning and memory ability of rats, while EGb761 and FPS-ZM1 may have certain protective effects on the nerve injury.
ABSTRACT
Objective: To evaluate the value of next-generation sequencing (NGS) in the prevention and management of thalassemia. Methods: A systematic search was performed in eight databases including China Biomedical Literature Database, Chinese National Knowledge Infrastructure, Chinese Scientific Journals Database, Wanfang database, PubMed, EMBASE, Web of Science, and Cochrane Library from the inception to 1 June 2022. Stata 17.0 and Review Manager 5.4 were used for the meta-analysis. Results: Nine studies containing 14794 participants were included in the meta-analysis. Compared with the routine genetic testing (including Gap-PCR and reverse dot blot), NGS had higher detection rates in screening thalassemia (RR 1.22, 95% CI 1.13-1.31, P0.05). Conclusions: Compared with routine genetic testing, NGS had a higher detection rate in general, particularly in the detection of α-thalassemia.
ABSTRACT
OBJECTIVE：To systematically evaluate clinical efficacy of L-carnitine in the treatment of male infertility ，and to provide evidence-based reference for clinical treatment of male infertility. METHODS ：Retrieved from CNKI ，Wanfang database ， VIP，CBM，PubMed，Embase and the Cochrane library ，randomized controlled trials （RCTs）about L-carnitine and other chemical drugs in the treatment of male infertility were collected during the inception to Apr. 12th，2020. After data extraction of included literatures and quality evaluation with modified Jadad scale ，Meta-analysis was conducted by using RevMan 5.3 software. RESULTS：A total of 8 RCTs were included ，with 520 patients. The results of Meta-analysis showed that compared with other chemical drugs ，L-carnitine could significantly enhance the semen volume [MD＝0.55，95％CI（0.20，0.91），P＝0.002] and sperm mortality rate [MD＝1.60，95％CI（0.50，2.69），P＝0.004] of male infertility patients ，with statistical significance. There was no statistical significance in sperm count [MD＝4.00，95％ CI（－3.15，11.15），P＝0.27]，the percentage of forward motile sperm [MD＝12.58，95％CI（－3.87，29.03），P＝0.13]，and the percentage of inducing pregnancy rate [OR＝0.85，95％CI（0.47，1.52），P＝ 0.58] of male infertility patients. CONCLUSIONS ：L-carnitine can significantly improve the semen volume and sperm mortality of male infertility patients ，and has the same effects as other drugs on improving sperm count ，percentage of forward motile sperm and percentage of inducing pregnancy rate.
ABSTRACT
BACKGROUND: Beta thalassaemia is a monogenic disease, which lacks effective clinical treatments. Hematopoieticstem cell transplantation currently is the only radical treatment for beta thalassaemia, but the limits of suitable donor and costs minimize its clinical application. Given the technology of reprogramming using somatic cells is well established,gene therapy using induced pluripotent stem cells has become the new direction of beta thalassaemia treatment.OBJECTIVE: To put forward the advantages of CRISPR/Cas9 technology in gene therapy of beta thalassaemia in thefuture by summarizing the mechanisms of three kinds of gene editing technologies and the preliminary experimentalresults in animal models.METHODS: In order to search relevant articles about beta thalassaemia, the first author retrieved PubMed database andCNKI (from 1989 to 2015) using the key words of beta thalassemia, genetic therapy, genome editing, homologousrecombination, iPSCs in English and Chinese, respectively. After eliminating literatures which were irrelevant toresearch purpose or containing a similar content, 67 articles were chosen for further analysis.RESULTS AND CONCLUSION: Gene editing technology has made considerable progress and three kinds of directedgene editing technologies have been developed, including ZFNs, TALENs, CRISPR/Cas technology. By targeting inducedpluripotent stem cells from thalassemi patients, these three kinds of gene editing technologies have been expected tocorrect pathogenic genes of thalassemia. The CRISPR/Cas system is more simple, rapid, safe and efficient than the others.The CRISPR/Cas9 system is expected to repair β-globin genes in the induced pluripotent stem cells, germ cells, fertilizedeggs and embryos from beta thalassaemia patients, laying the foundation for future clinical application.
ABSTRACT
BACKGROUND:Induced pluripotent stem cel technology have solved the contradiction between the ethics and immune rejection, and this high-efficient and safe technique is becoming the mainstream of today’s research. OBJECTIVE:To comprehensively review the safety and application of induced pluripotent stem cel s.METHODS:A computer-based online retrieval of PubMed and CNKI was performed to search relevant papers published from January 2006 to April 2016, with the key words of“induced pluripotent stem cel , reprogramming, clinical application, safety, transcription factor, disease mode”in English and Chinese, respectively. RESULTS AND CONCLUSION:In recent years, research on induced pluripotent stem cel s has attracted much attention from the scientific community and the medical community, and this technique has successful y gained induced pluripotent stem cel s and overcome the problems of immunity and ethics. However, it is limited to the theoretical and laboratory research due to the inability to solve the safety, efficiency and re-differentiation mechanism of induced pluripotent stem cel s. Therefore, we are faced with enormous difficulties and chal enges, which involve al aspects of basic research, including how to safely and effectively induce the differentiation of induced pluripotent stem cel s into the desired cel type and how to establish a suitable disease model as wel as a high-throughput drug screening platform.
ABSTRACT
Objective To investigate the influence of total progressively motile sperm count(TPMSC) after treatment on clinical outcomes of intrauterine insemination(IUI) with the husband′s sperm in ovulation-promoting cycles.Methods The clinical data in 4179 cases undergoing IUI with the husband′s sperm in ovulation-promoting cycles were retrospectively analyzed.The correlation between clinical pregnancy rate and TPMSC was analyzed.Results Among all the clinical data,TPMSC was to 100×106 in occasional live sperm.TPMSC60×106 had no pregnancy.A total of 4 154 cases of TPMSC (0.15-60.00)×106 were analyzed.The female age,duration of infertility,number of follicles and endometrial thickness(EDM) had no statistical differences among various groups.The clinical pregnancy rate was 13.5%(576/4 154),the group with the highest clinical pregnancy rate was (5.00-<10.00)×106.But there was no statistically significant difference in clinical pregnancy rate among groups(P=0.133).Conclusion Performing IUI in PMSC (0.15-60.00)×106 after processing can get preferable pregnancy rates.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the mechanism of miR-133a in reversing neonatal rat cardiomyocyte hypertrophy induced by phenylephrine.</p><p><b>METHODS</b>A miR-133a precursor cDNA was used to construct an adenovirus vector, which was transfected into 293 cells to harvest miR-133a-containing virus. Neonatal rat cardiac myocytes treated by phenylephrine were exposed to miR-133a adenovirus, and the changes in cell area was measured; the expression levels of miR-133a and Acta1, Actc1, Actb, Myh6, Myh7, and BNP mRNAs were detected by quantitative RT-PCR.</p><p><b>RESULTS</b>Phenylephrine treatment increased the area of cardiomyocytes by more than 3 folds and significantly enhanced the expression levels of Acta1, Actc1, Actb, Myh6, Myh7 and BNP mRNAs. All these changes were obviously reverse by miR-133a treatment.</p><p><b>CONCLUSION</b>miR-133a is an important regulator of phenylephrine-induced cardiomyocyte hypertrophy and negatively regulates this process.</p>
Subject(s)
Animals , Rats , Adenoviridae , Cells, Cultured , Genetic Vectors , Hypertrophy , MicroRNAs , Genetics , Myocytes, Cardiac , Cell Biology , Pathology , Phenylephrine , RNA, Messenger , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To estimate the value of blastocyst culture for preimplantation genetic diagnosis (PGD).</p><p><b>METHODS</b>Day 3 embryos were biopsied and analyzed with fluorescence in situ hybridization (FISH) technique. Embryos with normal FISH results were cultured into blastocysts, and the ones with better morphology scores were transferred. Fourteen embryos with abnormal FISH results were cultured into blastocysts. Part of the cells taken from the blastocysts were amplified by whole genomic amplification (WGA) and assessed by array-based comparative genomic hybridization (array-CGH) analysis.</p><p><b>RESULTS</b>Six blastocysts with normal FISH results were transferred in 5 cycles. Four healthy babies of 3 cycles were delivered. Another one was a singleton pregnancy but with embryo growth arrest, whose villus karyotype was normal. Fourteen embryos with abnormal FISH results were cultured into blastocysts and analyzed by array-CGH. Six blastocysts were normal by array-CGH.</p><p><b>CONCLUSION</b>FISH combined with blastocyst culture may further ensure the accuracy of PGD result. Detection at the blastocyst stage can avoid false positive results and mosaic interferences on Day 3 stage and are therefore more authentic.</p>
Subject(s)
Adult , Female , Humans , Male , Pregnancy , Blastocyst , Cell Biology , Comparative Genomic Hybridization , Methods , Embryo Transfer , Genetic Diseases, Inborn , Diagnosis , Embryology , Genetics , In Situ Hybridization, Fluorescence , Methods , Preimplantation Diagnosis , MethodsABSTRACT
BACKGROUND:At present, the development of reprogramming technology provides a wide prospect for stem cellresearch. Through the ectopic co-expression of reprogramming factors, the somatic cells can be reprogrammed to a pluripotent state, termed as induced pluripotent stem cells, which can avoid the ethical controversy faced in the research and application of embryonic stem cells. Also, we can generate patient-specific and disease-specific induced pluripotent stem cells, which significantly decrease immuno-rejection. However, reprogramming technology faces some chal enges, such as low efficiency and safety. OBJECTIVE:Based on the characteristics of induced pluripotent stem cells and the principles of reprogramming, to detail the progress in reprogramming technology from five aspects, including cellresources, carriers, transcription factors, microRNA and signal transduction pathway. METHODS:A computer-based online retrieval was performed to search papers published form January 1990 to April 2013 in VIP periodical ful-text database, Wanfang periodical ful-text database, CNKI periodical ful-text database, PubMed database and Springer database with key words of“reprogramming, induced pluripotent stem cell, signal transduction pathway, epigenetics, microRNA, transcription factor, vector, somatic cell, smal molecule compound, safety”both in Chinese and English. After excluding objective-independent papers, 67 papers were included for further analysis. RESULTS AND CONCLUSION:By exploring different cellresources, different carriers, various combination of transcription factors, microRNAs or inhibition of the signal transduction pathways, the reprogramming efficiency and safety have been improved greatly. However, currently, induced pluripotent stem cells stil could not meet the requirement of clinical application. To achieve the clinical application of induced pluripotent stem cells, it is urgent to explore the mechanism of reprogramming, and to optimize the programming strategy.
ABSTRACT
Objective This study investigated high fat diet influence on the changes of vitamin D receptor (VDR) expression and endothelial nitric oxide synthase (eNOS) in apolipoprotein E-deficient(apoE-/-) mice.MethodsApoE-/- mice and C57BLP6J mice were divide into two groups (normal control and high fat diet),high fat diet group were feed high fat feedstuff.Plasma 25-(OH)D levels were determined by competitive protein binding radioimmunity,VDR expression were determined by immunofluorescence and reverse transcription-polymerase chain reaction.The levels of NO and eNOS were determined by nitrate reductase.ResultsCompared with normal control group,high fat diet caused more severe dam-age of atherosclerosis in wild type mice and apoE-/- mice.In apoE-/- mice,the levels of plasma 25-(OH)D were significantly decreased [(26.44±1.28) ng/mL,(22.68±2.07)ng/mL,(17.46±2.22)ng/mL,(15.88±0.97)ng/mL,P＜0.01],the expression of VDR protein and mRNA were significantly increased[VDR :0.244±0.088,0.346±0.132,0.547±0.128,0.768±0.162;VDRmRNA:0.228±0.083,0.375±0.103,0.451±0.117,0.597±0.131,P＜0.01],and the levels of NO and eNOS were significantly increased[NO:(39.74±4.81)μmol/L,(48.1±5.24 )μmol/L,(67.34±6.14 )μmol/L,(86.74±8.05)μmol/L;eNOS:(8.6±0.77 )U/L,(12.28±1.42)U/L,(15.96±0.92)U/L,(18.68±1.15)U/L,P＜0.01].These changes were more significantly in high fat diet group(P＜0.01).ConclusionsThere were abnormalities of plasma 25-(OH)D level,VDR expression and the level of NO and eNOS in apoE-/- mice.These changes were more significantly in high fat diet group.
ABSTRACT
Coronaviruses are the causative agent of respiratory and enteric diseases in animals and humans. One example is SARS, which caused a worldwide health threat in 2003. In coronaviruses, the structural protein N (nucleocapsid protein) associates with the viral RNA to form the filamentous nucleocapsid and plays a crucial role in genome replication and transcription. The structure of N-terminal domain of MHV N protein also implicated its specific affinity with transcriptional regulatory sequence (TRS) RNA. Here we report the crystal structures of the two proteolytically resistant N- (NTD) and C-terminal (CTD) domains of the N protein from murine hepatitis virus (MHV). The structure of NTD in two different crystal forms was solved to 1.5 Å. The higher resolution provides more detailed structural information than previous reports, showing that the NTD structure from MHV shares a similar overall and topology structure with that of SARS-CoV and IBV, but varies in its potential surface, which indicates a possible difference in RNA-binding module. The structure of CTD was solved to 2.0-Å resolution and revealed a tightly intertwined dimer. This is consistent with analytical ultracentrifugation experiments, suggesting a dimeric assembly of the N protein. The similarity between the structures of these two domains from SARS-CoV, IBV and MHV corroborates a conserved mechanism of nucleocapsid formation for coronaviruses.
Subject(s)
Amino Acid Sequence , Binding Sites , Crystallography, X-Ray , Molecular Sequence Data , Murine hepatitis virus , Chemistry , Metabolism , Nucleocapsid Proteins , Chemistry , Metabolism , Phosphoproteins , Chemistry , Metabolism , Protein Binding , Protein Folding , Protein Multimerization , Protein Structure, Secondary , Protein Structure, Tertiary , RNA , Metabolism , Sequence AlignmentABSTRACT
Objective To establish the method to detect the expression of apolipoprotein E gene in human adipose cells. Methods Apolipoprotein E gene expression was determined by competitive reverse transcription-PCR in adipose cells of healthy children. Results Apolipoprotein E gene expressed in the adipose cells of healthy children. The mean expression quantity of apolipoprotein E mRNA in adipose cells was (0.41?0.12 ) in healthy children. Conclusions Competitive reverse transcription-PCR is a rapid, simple, sensitive and accurate method to analyze expression of apolipoprotein E gene.