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Ex vivo culture-amplified mesenchymal stem cells (MSCs) have been studied because of their capacity for healing tissue injury. MSC transplantation is a valid approach for promoting the repair of damaged tissues and replacement of lost cells or to safeguard surviving cells, but currently the efficiency of MSC transplantation is constrained by the extensive loss of MSCs during the short post-transplantation period. Hence, strategies to increase the efficacy of MSC treatment are urgently needed. Iron overload, reactive oxygen species deposition, and decreased antioxidant capacity suppress the proliferation and regeneration of MSCs, thereby hastening cell death. Notably, oxidative stress (OS) and deficient antioxidant defense induced by iron overload can result in ferroptosis. Ferroptosis may inhibit cell survival after MSC transplantation, thereby reducing clinical efficacy. In this review, we explore the role of ferroptosis in MSC performance. Given that little research has focused on ferroptosis in transplanted MSCs, further study is urgently needed to enhance the in vivo implantation, function, and duration of MSCs.
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Humans , Antioxidants/metabolism , Ferroptosis , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells , Iron Overload/metabolismABSTRACT
Objective:To explore the effect of blood flow restriction training on the quadriceps femoris and on knee stability after anterior cruciate ligament reconstruction.Methods:Forty patients recovering from anterior cruciate ligament reconstruction were randomly divided into an experimental group and a control group, each of 20. In addition to routine rehabilitation training, the control group was given routine knee flexion and extension strength training, while the experimental group trained for an additional 20 minutes doing knee flexion and extension resistance training with the blood pressure in their groins at 70% of their individual arterial occlusive pressure. (The mean pressure was (123±11.23)mmHg). The training lasted 8 weeks, three times a week. Knee function and hamstring and quadriceps peak torque were assessed before and after the intervention using a Lysholm scale and Humac Norm isokinetic muscle strength tests.Results:There were no significant differences between the two groups in any of the measurements before the training. After the intervention, all of the measurements had improved significantly in both groups, with the average Lysholm score, H/Q% and peak torque of the experimental group significantly better than the control group′s averages.Conclusions:Blood flow restriction training can improve the effectiveness of quadriceps femoris strength, knee stability and knee function training after anterior cruciate ligament reconstruction.
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Objective:To explore any effect of blood flow restriction during exercise on knee proprioception and motor coordination after anterior cruciate ligament (ACL) reconstruction.Methods:Thirty patients recovering from ACL reconstruction were randomly divided into an experimental group and a control group, each of 15. Both groups were given routine rehabilitation training, while the experimental group was additionally provided with 45 minutes of training with blood flow restriction, 3 times a week for 8 weeks. The blood flow restriction training involved constant pressure in the groin while the patient performed knee flexion and extension resistance training, squats, alternate knee flexion and extension and ergometer cycling. Before and after the intervention, both groups′ knee function, proprioception and lower limb motor coordination were evaluated using the Lysholm knee scale, the Humac isokinetic measurement system and surface electromyography.Results:Before the experiment there were no significant differences between the two groups in any of the measurements. After the intervention, both groups′ average Lysholm score had improved significantly, and errors in reproducing a knee angle had decreased significantly. Significantly better improvement was observed in the observation group than in the control group. That group′s average coordinated contraction rate on the affected side in extension and flexion was also significantly better than the control group′s ave-rage. Indeed, there were no significant differences in the contraction rates between the healthy and affected sides.Conclusions:Training with restricted blood flow can significantly improve knee function, proprioception and motor coordination after anterior cruciate ligament reconstruction.
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Objective:To evaluate the changes of retinal thickness, choroid thickness and choroid vascular polypoid lesions (polyps) before and after the treatment of conbercept in polypoidal choroidal vasculopathy (PCV) with conbercept based on enhanced-depth imaging optical coherence tomography (EDI-OCT) and OCT angiography (OCTA).Methods:Twenty patients with PCV admitted to the ophthalmology department of the Affiliated Hospital of Traditional Chinese Medicine of Xinjiang Medical University from January 2019 to December 2020 (who had not received intravitreous injection before) were selected and given intravitreous injection of conbercept with 3+ pro re nata (PRN) treatment. The changes of visual acuity, intraocular pressure, central macular thickness (CMT), subfoveal choroidal thickness (SFCT), peripapillary choroid thickness (PCT), polyps height, and maximum cross-sectional area of polyps were observed before and after treatment.Results:There were significant difference in the best corrected visual acuity (BCVA), CMT, SFCT, PCT, polyps height and maximum polyps cross-sectional area before and after treatment in 20 patients (all P<0.05). BCVA was significantly improved at 1 week and 1 month after treatment (all P<0.05), and still improved at 6 months after treatment, but the difference was not statistically significant compared with 3 months after treatment ( P>0.05), and slightly fluctuated at 6 to 12 months after treatment, but basically stable. CMT, SFCT and PCT decreased significantly after 3 months of treatment, and were basically stable from 6 to 12 months after treatment. One month after treatment, polyps height was (251.30±57.49)μm, with a decrease of 11.3 μm; the polyps height decreased at 3, 6 and 12 months after treatment, and the difference was statistically significant (all P<0.05). The maximum cross-sectional area of polyps at 3 and 12 months after treatment was (2.10±0.44)μm 2 and (1.98±0.44)μm 2, respectively, with a certain degree of decrease, and the difference was statistically significant (all P<0.05). Conclusions:Intraocular injection of conbercept can effectively reduce CMT, SFCT, PCT, polyps height, maximum cross-sectional area of polyps and increase BCVA in PCV patients 3 months after treatment. EDI-OCT and OCTA were used to quantitatively detect the changes of PCV indexes and observe the curative effect and prognosis.
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[This corrects the article DOI: 10.1016/j.apsb.2021.04.013.].
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Objective:To investigate the efficacy of floating acupuncture therapy in the treatment of chronic refractory pressure ulcers and its effects on serum levels of superoxide dismutase, malondialdehyde and insulin-like growth factor-1.Methods:A total of 60 patients with chronic refractory pressure ulcers who received treatment in The Second Hospital of Jiaxing from October 2019 to October 2021 were included in this study. They were randomly divided into observation and control groups, with 30 patients in each group. The patients in the control group were given routine treatment, while the patients in the observation group were given floating acupuncture therapy based on routine treatment. Clinical efficacy, wound healing rate after 7, 14 and 21 days of treatment, the time of emerging granulation tissue, the time of shedding rotten meat, and the time of wound healing, changes in serum levels of superoxide dismutase, malondialdehyde and insulin-like growth factor-1 after 21 days of treatment relative to those before treatment were compared between the two groups.Results:Total response rate in the observation group was significantly higher than that in the control group [90.0% (27/30) vs. 66.7% (20/30), χ2 = 4.81, P < 0.05]. Wound healing rate after 7, 14 and 21 days of treatment in the observation group was significantly higher than that in the control group ( t = -5.45, -7.77, -7.51, all P < 0.001). The time of emerging granulation tissue, the time of shedding rotten meat, and the time of wound healing in the observation group were significantly shorter than those in the control group ( t = 5.69, 9.26, 8.91, all P < 0.001). After 21 days of treatment, serum levels of superoxide dismutase, malondialdehyde and insulin-like growth factor-1 in the observation group were (96.52 ± 10.43) U/mL, (4.32 ± 0.78) μmol/L, (43.52 ± 8.29) μg/L, respectively, which were significantly different from (83.76 ± 6.81) U/mL, (5.48 ± 0.92) μmol/L, (36.78 ± 9.18) ng/mL, in the control group ( t = 5.61, 5.27, 2.98, all P < 0.05). Conclusion:Floating acupuncture therapy is effective in the treatment of refractory pressure ulcer. It can accelerate wound healing, increase serum superoxide dismutase and insulin-like growth factor-1 levels, and decrease serum malondialdehyde level.
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Objective:To study the expression level and clinical significance of minichromosome maintenance protein 3(MCM3) in human gastric cancer.Methods:The expression levels of MCM3 mRNA and protein in gastric cancer and adjacent normal tissues were analyzed using public databases such as TCGA, CCLE and HPA. The clinicopathological features of 69 patients with gastric cancer were retrospectively analyzed, and the expression levels of MCM3 protein in tumor tissues and adjacent normal tissues were detected by immunohistochemical staining, and the relationship between it and clinicopathological features was analyzed. The interaction network of MCM3 proteins was analyzed using the STRING database.Results:The expression levels of MCM3 mRNA and protein in gastric cancer tissues were significantly higher than those in adjacent normal tissues ( P<0.05), and its high expression was correlated with the size of gastric cancer tumors ( P<0.05). In gastric cancer tissue, the expression of MCM3 was correlated with the expression level of proliferating cell nuclear antigen (PCNA) ( R=0.61, P<0.01), and there may be a protein-protein interaction. Conclusions:MCM3 plays an important role in gastric cancer by interacting with PCNA, and it is expected to become a new diagnostic and therapeutic target.
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Objective:To study the safety and treatment outcomes of portal vein embolization (PVE) combined with lenvatinib plus an anti-programmed death-1(PD-1) antibody to treat patients with initially unreasectable hepatocellular carcinoma (uHCC).Methods:This study retrospectively analyzed the data of six patients with uHCC who received first-line combined systemic therapy with lenvatinib plus an anti-PD-1 antibody, and then underwent pre-hepatectomy PVE at the Department of Liver Surgery at Zhongshan Hospital, Fudan University from May 2019 to November 2020. All enrolled patients were males, aged (54.6±6.2) (ranged 46 to 63) years. Tumor response and liver volume were evaluated by medical imagings once every 2 months (±2 weeks) and evaluated using the Response Evaluation Criteria in Solid Tumours (version 1.1). Patients were followed-up by outpatient interviews or by phone calls to record their survival and tumor outcome status.Results:Three of the six enrolled patients had Barcelona Clinic Liver Cancer stage A and three had stage B disease. One patient achieved a partial response and five patients had stable diseases. The mean ± s. d. future liver remnant (FLR) percentage was (29.0±8.9) % before PVE and the combination therapy, and was (41.3±10.8) % before the last evaluation for liver surgery ( t=10.79, P<0.001). Hepatectomy was carried out in five patients, and one patient who failed to develop significant FLR hypertrophy did not undergo hepatectomy. Grade B post-hepatectomy liver failure and major postoperative complications (i.e. pleural effusion requiring additional percutaneous drainage) occurred in one patient. After a median post-operative follow-up of 4.5 (range: 1.0-12.3) months, all five patients were alive and were tumor free. Conclusion:PVE followed by hepatectomy is feasible in a uHCC patients receiving systemic therapy with lenvatinib and an anti-PD-1 antibody.
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Objective:To investigate the safety and dose of 4D template (real-time adjustable angle template) in the treatment of advanced malignant tumors with 125I seeds. Methods:98 patients with advanced malignant tumors admitted to Department of Thoracic Surgery of Shaanxi Provincial Tumor Hospital were treated with 4D template-navigated radioactive 125I seed implantation from June 2018 to December 2019. Preoperative TPS plan, intraoperative optimization, postoperative verification of immediate dose and postoperative evaluation of implantation dose were performed. The treatment results were observed. Results:All 98 patients completed the seed implantation. The implantation dose of GTV of implantation site receiving external irradiation was (12 489±414) cGy and the dose of no external irradiation was (15 036±514) cGy. V 100% was 84.7%-94.1%, and 88.2%-93.7%. The implantation dose of CTV was (7 450±621) cGy, and (9 080±761) cGy. The quality of dose implantation was evaluated as: excellent in 89 cases (91%, 89/98), good in 7 cases (7%, 7/98), fair in 2 cases (2%, 2/98), and poor in 0 case, respectively. The symptom relief rate of patients with pain was 92%(36/39). The 1-and 2-year local control rates were 61%, 36% and 82%, 54% in patients treated with and without external irradiation, respectively. The difference was statistically significant ( P=0.02). The incidence rates of pneumothorax and hemoptysis were 19%(9/48) and 10%(5/48). No corresponding complications were observed in other parts of the patients. Conclusion:4D template-assisted 125I seed therapy is safe and effective for malignant tumors, and intraoperative adjustment of needle angle and dose optimization can realize the precise control of implantation dose.
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As a structural barrier, the blood-brain barrier (BBB) is located at the interface between the brain parenchyma and blood, and modulates communication between the brain and blood microenvironment to maintain homeostasis. The BBB is composed of endothelial cells, basement membrane, pericytes, and astrocytic end feet. BBB impairment is a distinguishing and pathogenic factor in diabetic encephalopathy. Diabetes causes leakage of the BBB through downregulation of tight junction proteins, resulting in impaired functioning of endothelial cells, pericytes, astrocytes, microglia, nerve/glial antigen 2-glia, and oligodendrocytes. However, the temporal regulation, mechanisms of molecular and signaling pathways, and consequences of BBB impairment in diabetes are not well understood. Consequently, the efficacy of therapies diabetes targeting BBB leakage still lags behind the requirements. This review summarizes the recent research on the effects of diabetes on BBB composition and the potential roles of glial and vascular cells as therapeutic targets for BBB disruption in diabetic encephalopathy.
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Objective:To analyze the phenotype and genotype of two propositi with inherited hypodysfibrinogenaemia caused by compound heterozygous mutations, and investigate the molecular mechanism.Metheds:Two propositi and their family members(7 person in 3 generations and 10 person in 3 generations,respectively) were investigated. The activity of plasma fibrinogen (Fg:C) and thrombin time (TT) were analyzed by coagulation method, the antigen of plasma fibrinogen (Fg:Ag) was detected by immunoturbidimetry. All of the exons and flanking sequences of FGA,FGB,FGG of two propositi were amplified by PCR, followed by direct sequencing. The ClustalX-2, 1-win software was used to analyze the conservatism of mutated gene locus. PROVEAN and Mutation Taster were applied to analyze the pathogenicity of mutated amino acid. The changes of the protein spatial structure and intermolecular interaction were analyzed by Pymol.Results:Fg:C and Fg:Ag of proposita A and B were both significantly decreased (0.74 and 0.78 g/L, 0.96 and 0.94 g/L, respectively). Gene analysis revealed that proposita A and B both carried a heterozygous mutation c.2185G>A(p.AαGlu710Lys) in exon 6 of FGA. Furthermore, proposita A also carried a heterozygous mutation c.701G>T(p.γTrp208Leu) in exon 7 of FGG, and proposita B carried a heterozygous mutation c.1015A>C(p.γSer313Arg) in exon 8 of FGG. Phylogenetic analysis suggested that p.AαGlu710,p.γTrp208 and p.γSer313 were highly conserved among homologous species. All variants were predicted to be deleterious by two online bioinformatic softwares. The protein model analysis indicated that protein spatial structure and intermolecular hydrogen bonds were changed by these variants, which destroyed the stability of Fg.Conclusion:The compound heterozygous mutations of p.AαGlu710Lys and p.γTrp208Leu,p.AαGlu710Lys and p.γSer313Arg might account for the hypodysfibrinogenemia in two propositi.
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Objective@#To analyze delay in student pulmonary tuberculosis(PTB) case finding and associated factors in Suzhou, and to provide a reference for tuberculosis outbreak prevention and control in schools.@*Methods@#A total of 1 148 students with PTB who registered and were treated in Suzhou from 2011 to 2020 were included. Kruskal Wallis H test, 2 test and Cochran Armitage trend test were used to analyze the time trend of case finding delay. Logistic regression was used to analyze the correlation between admission characteristics and case finding delay.@*Results@#Among the students with PTB, a total of 569 cases were found to be delayed. The rate of delay was 49.6%, and the median delay time was 26(11-49) days. From 2011 to 2020, the difference in case finding interval of students with PTB was statistically significant( Hc=54.62, P <0.05), and the difference in case finding rate was also statistically significant( χ 2=53.69, P <0.05). The rate of delay fluctuated, with an overall upward trend over time( Z=-3.67, P < 0.05). Clinical consultation( OR=5.57, 95%CI =1.91-16.27), positive etiology ( OR=1.46, 95%CI =1.14-1.86) were positively correlated with case finding delay(all P <0.05).@*Conclusion@#There are significant delays in case finding among students with PTB in Suzhou. Clinical consultation and positive etiology are associated with case finding delay. In response to the growing problems in daily school tuberculosis prevention and control, multiple departments should cooperate to implement relevant measures and to reduce the occurrence of case finding delay.
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OBJECTIVE：To study the improvement effects of paeon iflorin（PF）on myocardial injury in type 2 diabetes mellitus（T2DM）model rats and its mechanism. METHODS ：The experiment was set up in the normal group ，model group ， positive control group （metformin 90 mg/kg），PF high-dose ，medium-dose and low-dose groups （90，60，30 mg/kg），with 8 rats in each group. Except for normal group ，other groups were given high-glucose and high-fat diet and intraperitoneal injection of streptozotocin （30 mg/kg） to induce T 2DM model. After modeling ， administration groups were given relevant medicine intragastrically，normal group and model group were given constant volume of normal saline intragastrically ，once a day ，for consecutive 4 weeks. The body weight ，fasting blood glucose and oral glucose tolerance were measured ；serum levels of glycosylated serum protein （GSP），total cholesterol （TC），triacylglycerol（TG），glutathione peroxidase （GSH-Px），superoxide dismutase（SOD），malondialdehyde（MDA），creatine kinase isoenzyme-MB （CK-MB） and troponin Ⅰ （cTn Ⅰ） were determined. The pathomorphological changes of myocardium were observed. The apoptosis index of rat cardiomyocytes was （ detected. The protein expression of B-cell lymphoma 2 （Bcl-2），Bcl-2 related X protein （Bax）and caspase- 3 in rat myocardium were detected by immunohistochemistry and Western blot. RE SULTS：Compared with normal group ，the body weight ，serum levels of GSH-Px and SOD ，protein expression of Bcl- 2 in myocardium were decreased significantly in model group（P＜0.01）；while fasting blood glucose ，area under blood glucose curve ，serum levels of biochemical indexes （GSP，TC， TG，MDA，CK-MB，cTnⅠ），cardiomyocyte apoptosis index ，protein expression of Bax and caspase- 3 in myocardium were increased significantly （P＜0.05 or P＜0.01）. The arrangement of myocardium was relatively irregular ，and some muscle fibers were broken. Compared with model group ，except for body weight ，serum levels of SOD and MDA ，the protein expression of Bax in myocardium in PF low-dose group ， above indexes of PF groups were reversed significantly （P＜0.05 or P＜0.01）. CONCLUSIONS：PF can regulate glycolipid metabolism ，enhance antioxidant ability ，inhibit cardiomyocyte apoptosis and improve myocardial injury in T 2DM model rats ；the mechanism may be associated with increasing the protein expression of Bcl- 2 and down-regulating the protein expression of Bax and caspase- 3 in myocardium.
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Vascular smooth muscle cell (VSMC) migration plays a critical role in the pathogenesis of many cardiovascular diseases. We recently showed that TMEM16A is involved in hypertension-induced cerebrovascular remodeling. However, it is unclear whether this effect is related to the regulation of VSMC migration. Here, we investigated whether and how TMEM16A contributes to migration in basilar artery smooth muscle cells (BASMCs). We observed that AngII increased the migration of cultured BASMCs, which was markedly inhibited by overexpression of TMEM16A. TMEM16A overexpression inhibited AngII-induced RhoA/ROCK2 activation, and myosin light chain phosphatase (MLCP) and myosin light chain (MLC20) phosphorylation. But AngII-induced myosin light chain kinase (MLCK) activation was not affected by TMEM16A. Furthermore, a suppressed activation of integrin
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Objective:To analyze the expressions of minichromosome maintenance protein 4(MCM4) and proliferating cell nuclear antigen (PCNA) in gastric cancer tissues, to explore the relationship between MCM4/PCNA and gastric cancer, and to investigate the possibility of MCM4/PCNA as a potential biomarker for gastric cancer.Methods:Bioinformatics methods were used to analyze the mRNA expressions of MCM4 and PCNA in gastric cancer tissues and adjacent normal tissues. The clinicopathological data of 69 patients with gastric cancer who underwent surgery were retrospectively analyzed. The expression levels of MCM4 and PCNA in gastric cancer tissues and adjacent normal tissues were detected by immunohistochemistry, and their relationship with the clinicopathological characteristics of gastric cancer patients was analyzed.Results:The mRNA levels of MCM4 and PCNA in gastric cancer tissues are significantly higher than those in adjacent normal tissues (all P<0.05). The expression of MCM4 is correlated with the tumor size of gastric cancer ( P=0.037), but there is no significant correlation with gender, age and tumor grade (all P>0.05). Both MCM4 and PCNA proteins are highly expressed in gastric cancer patients. Conclusions:The expression levels of MCM4 and PCNA have a clear correlation with the occurrence of gastric cancer. MCM4 and PCNA are expected to be potential biomarkers for the diagnosis and treatment of gastric cancer.
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OBJECTIVE@#To analyze the phenotype and genetic variants of a pedigree affected with hereditary protein C (PC) deficiency.@*METHODS@#The protein C activity (PC:A) of the proband and her family members (a four-generation pedigree including 11 individuals) were tested by chromogenic substrate method, and the protein C antigen (PC:Ag) was detected with an enzyme-linked immunosorbent assay(ELISA). The 9 exons and flanking sequences of the protein C (PROC) gene were amplified by PCR and directly sequenced. Suspected mutation was validated by clone sequencing and in other members of the family. MutationTaster and ClustalX-2.1-win was used to analyze the pathogenicity and conservation of the mutation site,respectively. Three-dimentional protein model and amino acids interaction were analyzed with Swiss-PdbViewer software.@*RESULTS@#The PC: A and PC: Ag of the proband were decreased to 46% (reference range: 70%-130%) and 50% (referencerange:70%-140%), respectively. Her grandmother,aunt, cousin and younger brother also showed declined PC:A and PC:Ag by approximately 50%. Genetic analysis revealed that the above individuals have all carried a deletional mutation c.1212-1212delG (p.Met364TrpfsX15) in exon 9 of the PROC gene which can cause replacement of Methionine at position 364 by Tryptophan and alteration of 15 downstream amino acids, and produce a premature stop codon at position 378. The score of MutationTaster was 1.000, indicating that the variant is pathogenic. Conservative analysis showed that the 15 altered amino acids are located in a conserved region across nine homologous species. Protein model analysis showed that the mutation has disrupted a catalytic domain of protein C thereby affected its function.@*CONCLUSION@#The heterozygous c.1212-1212delG deletional mutation in exon 9 of the PROC gene, which was unreported previously,probably accounts for the decrease of PC:A and PC:Ag in this pedigree.
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OBJECTIVE@#To analyze the phenotype and genetic basis for a pedigree affected with hereditary coagulation factor XI deficiency.@*METHODS@#Activated partial thromboplastin time (APTT), prothrombin time (PT), fibrinogen (FIB), FXI activity (FXI:C) and the antigen of FXI (FXI:Ag) were determined for the proband and members from his pedigree. Sanger sequencing was used to analyze all exons, exon-intronic boundaries, as well as the 5'- and 3'- untranslated regions of the F11 gene. Suspected variants were verified in her family members and confirmed by reverse sequencing. The impact of the variants on the protein function was predicted by using PolyPhen-2 and SIFT software. The protein structure and amino acid interaction were analyzed by using Swiss-PdbViewer.@*RESULTS@#The APTT, FXI:C and FXI:Ag of the proband and her sister were significantly reduced to 73.0 s, 10.0%, 15.0% and 87.1 s, 2.0% and 11.5%, respectively. APTT of some family members was slightly prolonged, and FXI:C and FXI:Ag also decreased to various extents. DNA sequencing revealed that the proband and her sister have carried compound heterozygous variants of c.738G>A (p.Trp228stop) and c.938G>T (p.Ser295Ile) respectively in exons 7 and 9 of the F11 gene. Her father, sister and daughter were heterozygous for the c.738G>A (p.Trp228stop) variant, while her mother and nephew were heterozygous for the c.938G>T (p.Ser295Ile). Both PolyPhen-2 and SIFT predicted that the p.Ser295Ile variant is likely to be deleterious and can affect the protein function. Modeling analysis indicated that the p.Ser295Ile variant may lead to disruption of a hydrogen bond, resulting in alteration of protein structure and instability.@*CONCLUSION@#The compound heterozygous c.738G>A (p.Trp228stop) and c.938G>T (p.Ser295Ile) variants of the F11 gene probably underlie the decreased FXI level in this pedigree.
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Female , Humans , Factor XI Deficiency , Genetics , Genetic Variation , Heterozygote , Mutation , PedigreeABSTRACT
OBJECTIVE@#To explore the molecular basis for a Chinese pedigree affected with hereditary coagulation factor VII (FVII) deficiency.@*METHODS@#The coding regions of F7 gene were amplified by PCR and sequenced. Suspected variants were confirmed by reverse sequencing and validated in other members from the pedigree. Pathogenicity of the variants was analyzed with multiple bioinformatic tools.@*RESULTS@#Genetic analysis revealed that the proband has carried compound heterozygous c.985T>C (p.Ser329Pro) and c.1091G>A (p.Arg364Gln) variants in exon 8 of the F7 gene. Her mother, brother and son were heterozygous for c.985T>C (p.Ser329Pro), while her father was heterozygous for c.1091G>A (p.Arg364Gln). Phylogenetic analysis suggested that both p.Ser329 and p.Arg364 are highly conserved among homologous species. Online bioinformatic software predicted both variants to be deleterious. Protein model analysis suggested that the Pro329 side chain may form a new hydrogen bond with Leu333. The Pro benzene ring may clash with Glu325 in the p.Ser329Pro variant model. The p.Arg364Gln variant have two additional hydrogen bonds compared with wild type Arg364. Both variants may lead to alteration of the protein structure.@*CONCLUSION@#The p.Ser329Pro and p.Arg364Gln variants in exon 8 of the F7 gene probably account for the reduced FVII in this pedigree.
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Objective: To summarize the research progress of the regulatory role of microRNA (miRNA) in osteogenic differentiation of mesenchymal stem cells (MSCs) and its application as a therapeutic target and diagnostic tool in orthopedic diseases. Methods: The recent literature on the regulation of MSCs osteogenic differentiation by miRNAs was extensively reviewed, and its regulatory mechanism and its application as a therapeutic target and diagnostic tool in orthopedic diseases were reviewed. Results: miRNAs are small endogenous non-coding RNAs with a length of 20-22 nucleotides, which play an important role in the osteogenic differentiation of MSCs. Osteogenesis begins with the differentiation of MSCs into mature osteoblasts, and each stage of dynamic homeostasis of bone metabolism is associated with the regulation of different miRNAs. miRNAs are regulated from the post-transcriptional level by mRNAs cleavage, degradation, translational repression, or methylation. In addition, current studies suggest that miRNAs can be used as a new diagnostic tool and therapeutic target for orthopedic diseases. Conclusion: Further study on the regulation mechanism of miRNAs will provide more ideas for finding new therapeutic targets and diagnostic tools for orthopedic disease.
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Objective@#To detect potential mutations of the coagulation factor Ⅶ (F7) gene in a pedigree affected with hereditary FⅦ deficiency and explore its molecular pathogenesis.@*Methods@#The FⅦ antigen (FⅦ∶Ag) was analyzed by an enzyme-linked immunosorbent assay (ELISA) method. Prothrombin time (PT), FⅦ activity (FⅦ∶C) and other coagulant parameters were quantified with an one-stage clotting assay. The F7 gene was amplified by PCR and sequenced. Mutational sites were confirmed by reverse sequencing. Impact of amino acid substitution was assessed using SIFT and PolyPhen-2 software. Structure of the mutant protein was analyzed using Swiss-pdb Viewer software based on the three-dimensional structure in the Protein Data Bank.@*Results@#The propositus had prolonged PT (36.3 s), with FⅦ∶C and FⅦ∶Ag significantly reduced to 2% and 44%, respectively. Her father, mother, younger sister and daughter had slightly prolonged PT and reduced FⅦ∶C (86%-120%). The FⅦ∶Ag of her father and younger sister were also reduced. DNA sequencing revealed that the propositus has carried compound heterozygous mutations (Lys341Glu and IVS6-1G>A) of the F7 gene. Her father and younger sister were heterozygous for the IVS6-1G>A mutation, while her mother and daughter were heterozygous for the Lys341Glu mutation. Bioinformatics analysis indicated that Lys341Glu mutation may affect the stability and function of the FⅦ protein.@*Conclusion@#The Lys341Glu and IVS6-1G>A mutations probably underlie the reduced activity of FⅦ in this pedigree.