Single-nucleus transcriptomics reveals a gatekeeper role for FOXP1 in primate cardiac aging

Aging poses a major risk factor for cardiovascular diseases, the leading cause of death in the aged population. However, the cell type-specific changes underlying cardiac aging are far from being clear. Here, we performed single-nucleus RNA-sequencing analysis of left ventricles from young and aged cynomolgus monkeys to define cell composition changes and transcriptomic alterations across different cell types associated with age. We found that aged cardiomyocytes underwent a dramatic loss in cell numbers and profound fluctuations in transcriptional profiles. Via transcription regulatory network analysis, we identified FOXP1, a core transcription factor in organ development, as a key downregulated factor in aged cardiomyocytes, concomitant with the dysregulation of FOXP1 target genes associated with heart function and cardiac diseases. Consistently, the deficiency of FOXP1 led to hypertrophic and senescent phenotypes in human embryonic stem cell-derived cardiomyocytes. Altogether, our findings depict the cellular and molecular landscape of ventricular aging at the single-cell resolution, and identify drivers for primate cardiac aging and potential targets for intervention against cardiac aging and associated diseases.

4E-BP1 counteracts human mesenchymal stem cell senescence via maintaining mitochondrial homeostasis

Although the mTOR-4E-BP1 signaling pathway is implicated in aging and aging-related disorders, the role of 4E-BP1 in regulating human stem cell homeostasis remains largely unknown. Here, we report that the expression of 4E-BP1 decreases along with the senescence of human mesenchymal stem cells (hMSCs). Genetic inactivation of 4E-BP1 in hMSCs compromises mitochondrial respiration, increases mitochondrial reactive oxygen species (ROS) production, and accelerates cellular senescence. Mechanistically, the absence of 4E-BP1 destabilizes proteins in mitochondrial respiration complexes, especially several key subunits of complex III including UQCRC2. Ectopic expression of 4E-BP1 attenuates mitochondrial abnormalities and alleviates cellular senescence in 4E-BP1-deficient hMSCs as well as in physiologically aged hMSCs. These f indings together demonstrate that 4E-BP1 functions as a geroprotector to mitigate human stem cell senescence and maintain mitochondrial homeostasis, particularly for the mitochondrial respiration complex III, thus providing a new potential target to counteract human stem cell senescence.

Research of early outcomes of cementless Oxford unicompartmental knee arthroplasty / 中华骨科杂志

Objective:To assess the early clinical outcomes of Oxford cementless unicompartmental knee arthroplasty (UKA) in the management of anteromedial osteoarthritis of the anterior medial knee (AMOA).Methods:A retrospective review was conducted on a cohort of 90 patients who underwent primary UKA for AMOA at the Fuzhou Second Hospital between January 2020 and June 2021. The patients were divided into two groups based on the type of prosthesis used: the cementless UKA group and the cemented UKA group. The cementless UKA group included 45 patients (22 males and 23 females), with a mean age of 65.6±7.0 years (ranging from 52 to 81 years). The cemented UKA group consisted of 45 patients, including 21 males and 24 females, with a mean age of 67.9±6.1 years (ranging from 55 to 79 years). The study compared various parameters between the two groups, including the duration of surgery, amount of blood loss, length of hospitalization, pain visual analogue scale (VAS) scores, Hospital for Special Surgery (HSS) scores, and forgotten joint score (FJS). Additionally, radiolucency around the prosthesis, medial proximal tibial angle (MPTA), and mechanical axis deviation (MAD) were observed and measured before and after surgery to assess prosthesis stability, improvement of knee deformity, and knee joint mobility, respectively.Results:Both groups were followed up for a minimum of 18 months. No statistically significant differences were found in demographic data such as gender, age, and body mass index between the two groups. The cementless UKA group demonstrated superior results in terms of shorter duration of surgery (97.8±21.1 min) and less blood loss (70.8±37.6 ml) compared to the cemented UKA group ( P<0.05). However, no significant difference in length of stay was observed between the groups. Postoperative VAS scores, HSS scores, MPTA, and MAD were significantly improved compared to preoperative values in both groups ( P<0.05). The median VAS scores for the cementless UKA group at 6, 12, and 18 months postoperatively were 4.0(3.0, 5.0), 2.0(2.0, 3.0), and 2.0(1.0, 3.0) respectively. The corresponding median HSS scores for the cemented UKA group were 78(75, 82), 85 (80, 89), and 86(82, 90) preoperatively and at 6, 12, and 18 months postoperatively, respectively. The median FJS scores for the cementless UKA group were 73.0(70.5, 76.0), 76.0(74.0, 78.0), and 66.0(63.0, 68.0) preoperatively and at 6, 12, and 18 months postoperatively, respectively. There was no statistically significant relationship between preoperative VAS scores and preoperative FJS scores ( P>0.05). However, there was a significant difference in VAS scores between the two groups at 6 months postoperatively ( P<0.05). The cementless UKA group consistently exhibited better VAS scores and HSS scores compared to the cemented UKA group at 6, 12, and 18 months postoperatively ( P<0.05). The median MPTA values for the cementless UKA group were 85.41°(84.22°, 86.54°), 85.80°(84.74°, 87.41°), 86.51°(85.47°, 88.14°), and 86.80°(86.09°, 88.27°) preoperatively and at 6, 12, and 18 months postoperatively, respectively. The median MAD values for the cementless UKA group were 2.29(1.79, 2.65) cm, 1.11(0.69, 1.75) cm, 1.02(0.65, 1.66) cm, and 0.91(0.61, 1.63) cm preoperatively and at 6, 12, and 18 months postoperatively, respectively. There were no significant differences in MPTA and MAD between the two groups at all time points, except for the preoperative MPTA and postoperative MAD at 6 and 12 months. The incidence of physiological radiolucency around the prosthesis was significantly lower in the cementless UKA group 3 than in the cemented UKA group 28 ( P<0.05). However, no complete radiolucency was observed in either group. Conclusion:Oxford cementless UKA is a highly effective treatment for relieving joint pain and correcting knee deformity in patients with knee AMOA. It offers the advantages of shorter surgical duration, reduced blood loss, and improved subjective outcomes for patients after the operation.

Analysis of genetic variants in a pedigree affected with hereditary multiple osteochondroma / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a pedigree affected with hereditary multiple osteochondroma (HMO).@*METHODS@#Peripheral blood samples were collected from the proband and members of his pedigree with informed consent. Following extraction of genomic DNA, all coding exons and flanking intronic sequences (-10 bp) of the EXT1 and EXT2 genes were subjected to targeted capture and next generation sequencing (NGS). Suspected variant was verified by Sanger sequencing.@*RESULTS@#A heterozygous nonsense variant (c.1911C>A) was found in exon 10 of the EXT1 gene in the proband and his affected father but not in a healthy sister and normal controls. The variant was classified as a pathogenic based on the guidelines of the American College of Medical Genetics and Genomics (PVS1+PM2+PP1). Bioinformatic analysis predicted that the c.1911C>A variant may be disease-causing via nonsense-mediated mRNA decay and anomalous splicing.@*CONCLUSION@#The c.1911C>A variant probably underlay the disease in this pedigree. Discovery of this variant enriched the variant spectrum of HMO.

Application of phage in patients with urinary tract pandrug-resistant Klebsiella pneumoniae infection / 中华泌尿外科杂志

Objective:To summarize the application of phage therapy in patients with urinary tract complicated pandrug-resistant Klebsiella pneumoniae infection, and analyze its feasibility and effectiveness.Methods:To retrospectively analyze the clinical data of a patient with complicated urinary tract complex pan-resistant Klebsiella pneumoniae treated by phage from August to September, 2019 in Shanghai Public Health Clinical Center. The female patient, 65 years old, was admitted to the hospital on August 6, 2020. The patient repeated with frequent micturition and urgent micturition half a year before admission. These symptoms were not accompanied by back pain, fever, chills, dysuria, gross hematuria. Urinary culture results in outpatient hospital was pan-resistant Klebsiella pneumoniae. After the patient discontinued application of cefoperazone sulbactam, levofloxacin and other drugs, symptoms such as frequent urination could be relieved after treatment, but appeared repeatedly. In August 2019, the center innovatively applied phage therapy to treat this patient with urinary tract pandrug-resistant bacteria infection.Results:For the first time, we applied 117, 135, 178, GD168 phage mixed solution once a day, for 5 days of continuous bladder infusion. At the same time, meropenem and amikacin was intravenous administration to strengthened the anti-infection treatment. Urine culture was negative for two consecutive times after treatment. However, half a month after the end of the bladder infusion, the patient experienced discomfort such as frequent urination. Urine culture: pan-resistant Klebsiella pneumoniae. The second time, we applied a mixture of three phage strains 130, 131, 909, once a day, for 5 days of continuous bladder infusion. And in the afternoon of the third day of treatment, the renal pelvis was retrogradely intubated and perfused with the above three strains of phage mixture. During the second treatment follow-up until March 30, 2020, the patient's urine culture was reviewed once a month. As a result, no pan-resistant Klebsiella pneumoniae was found, and the patient no longer experienced frequent urination and other symptoms of urination. The treatment process was successful and without severe complications and side effects.Conclusions:Phage urinary tract perfusion is an effective method for the treatment of pan-resistant Klebsiella pneumoniae urinary tract infections. The curative effect is accurate and reliable. The patient did not show obvious complications and adverse reactions during treatment. It can be used as an alternative treatment plan for complex pan-resistant Klebsiella pneumoniae infection.

Fixed-bearing unicompartmental knee arthroplasty for anteromedial knee osteoarthritis or spontaneous osteonecrosis of the femoral condyle / 中华创伤骨科杂志

Objective:To investigate the clinical efficacy of fixed-bearing unicompartmental knee arthroplasty (UKA) in the treatment of anteromedial knee osteoarthritis or spontaneous osteonecrosis of the femoral condyle.Methods:Between June 2017 and June 2018, 62 consecutive patients were treated at Department of Joint Surgery, Fuzhou Second Hospital Affiliated to Xiamen University for anteromedial knee osteoarthritis or spontaneous osteonecrosis of the femoral condyle using a fixed-bearing UKA system. They were 27 men and 35 women, aged from 58 to 85 years (average, 69.8 years). There were 58 cases of anteromedial knee osteoarthritis and 4 cases of spontaneous osteonecrosis of the femoral condyle. The courses of disease ranged from 2 to 10 years (average, 5.1 years). Medial unicompartmental knee arthroplasty was performed for all the patients on the fixation platform SLED. Recorded were the hip-knee-ankle angle (HKA) of the patients 2 days after operation, Oxford knee score (OKS) and American Knee Society score (KSS) at the final follow-up and post-operative complications.Results:The 62 patients were followed up for 9 to 21 months (average, 15.6 months). Tibial plateau fracture occurred in one case who eventually obtained bony union after open reduction and internal fixation. Another case had a wound healing problem which responded to drainage and debridement with prosthesis preservation. Another case presented prepatellar pain which was improved after physical therapy. In the 62 patients, the HKA 2 days after operation (172.27°±1.61°) was significantly improved compared to the preoperative value (177.79°±1.32°), the OKS at the final follow-up (19.8±2.7) significantly lower than the preoperative value (50.2±3.1), and the KSS at the final follow-up (89.7±2.6) significantly higher than the preoperative value (49.6±5.2) (all P<0.05). Follow-ups revealed no cases of joint infection or prosthetic loosening. Conclusion:Fixed-bearing unicompartmental knee arthroplasty can lead to fine short-term outcomes for anteromedial knee osteoarthritis or spontaneous osteonecrosis of the femoral condyle.

Whether direct anterior approach for total hip arthroplasty is a risk factor of eccentric reaming to the anterior column of the acetabulum? / 中国组织工程研究

BACKGROUND:Direct anterior approach has been reported to be associated with a risk of reaming the anterior column of the acetabulum,but there is little evidence supporting this opinion.OBJECTIVE:To reveal differences in the bone stock of the anterior and posterior columns between the direct anterior approach and the posterolateral approach in total hip arthroplasty by CT measurement.METHODS:Sixty cases of primary total hip arthroplasty through direct anterior approach (n=30) or posterolateral approach (n=30) at Fuzhou Second Hospital of Xiamen University from October 2015 to December 2017 were enrolled.The cross-sectional area of the anterior and posterior column of the acetabulum,the height of the anterior and posterior column,acetabular diameter,and anteversion were measured by CT.All researchers had 5-10 years of clinical experience,and the surgeons were associate chief physicians or above.The trial has been approved by the Ethics Committee of Fuzhou Second Hospital of Xiamen University on June 1,2017.All patients signed the written informed consents.RESULTS AND CONCLUSION:(1) The postoperative cross-sectional area of the anterior and posterior column of the acetabulum,and the height of the anterior and posterior column in the two groups were less than those at baseline (P ＜ 0.05),the acetabular diameter was larger than that at baseline (P ＜ 0.01),and the anteversion showed no significant difference (P ＞ 0.05).The area of the anterior column in the direct anterior approach group was higher than that in the posterolateral approach group (P ＜ 0.05),and other parameters were insignificantly different between two groups (P ＞ 0.05).(2) in summary,compared with posterolateral approach in total hip arthroplasty,direct anterior approach leads to increased area of the anterior column,and made no effect on other parameters.Thereafter,direct anterior approach is not a risk factor for eccentric reaming the anterior column of acetabulum,and the conclusion needs to be confirmed by multicenter,prospective randomized controlled trials.

Telomere-dependent and telomere-independent roles of RAP1 in regulating human stem cell homeostasis

RAP1 is a well-known telomere-binding protein, but its functions in human stem cells have remained unclear. Here we generated RAP1-deficient human embryonic stem cells (hESCs) by using CRISPR/Cas9 technique and obtained RAP1-deficient human mesenchymal stem cells (hMSCs) and neural stem cells (hNSCs) via directed differentiation. In both hMSCs and hNSCs, RAP1 not only negatively regulated telomere length but also acted as a transcriptional regulator of RELN by tuning the methylation status of its gene promoter. RAP1 deficiency enhanced self-renewal and delayed senescence in hMSCs, but not in hNSCs, suggesting complicated lineage-specific effects of RAP1 in adult stem cells. Altogether, these results demonstrate for the first time that RAP1 plays both telomeric and nontelomeric roles in regulating human stem cell homeostasis.

Comparison of several common detection indexes of cell senescence / 中国药理学与毒理学杂志

Cell senescence is a state of irreversible growth arrest, which can be triggered by a variety of different cellular stress. Currently, the detection indexes involved in the study of cell senes?cence include senescence-associated β-galactosidase, telomeres and telomerase, senescence-associ?ated heterochromatin foci, senescence-associated secretory phenotype, reactive oxygen species, and tumor suppressor genes p53 and p16. These indexes are widely used in the study of cell senescence, each with its own characteristics or advantages. This review summarizes several common cell senes?cence indexes and compares their accuracy, credibility, specificity, and potential applications.

A patient with pseudoachondroplasia undergoing bilateral total knee arthroplasties using three-dimensional printing technology / 中国组织工程研究

BACKGROUND: Pseudoachondroplasia is rare heredigary osteopathy, usually accompanied by deformity of the lower limbs, short fingers, and ligamentous laxity, so it is a difficulty in total knee arthroplasty.OBJECTIVE: To evaluate the curative effectiveness of bilateral total knee arthroplasties for a patient with pseudoachondroplasia using three-dimensional (3D) printing technology.METHODS: A retrospective study was done in a case of pseudoachondroplasia undergoing bilateral total knee arthroplasties using 3D printing technology. X-ray examination was conducted to observe the prosthesis location after arthroplasties, and the Knee Society Score outcome measures were used to evaluate the functional outcome.RESULTS AND CONCLUSION: (1) After arthroplasties, X-rays showed good line of force of the lower limbs and prosthesis location, and 1 year later, showed no radiolucent lines in tibia. (2) The right Knee Society Score improved to 94 points, the functional scoring improved to 90 points, and the postoperative flexion was 95°; the left Knee Society Score improved to 93 points, the functional scoring improved to 90 points, and the postoperative flexion was 90°. All patients were very satisfied with the treatment outcomes. (3) These results indicate that the individualized treatment scheme designed using 3D printing technology can reduce the surgical difficulty and trauma with high accuracy, and promote the functional recovery of the knee. Additionally, it obtains good clinical efficacy and patient's satisfaction in orthopaedic surgeries.

Program optimization before enucleation on ovine somatic cell nuclear transfer / 生物工程学报

Ovine somatic cell nuclear transfer (SCNT) efficiency remains lower. Therefore, we optimized the program before oocyte enucleation on ovine SCNT. Four experiments were done including exposure duration of ovaries (3 h or 3 to 5 h), duration of oocytes maturation (18 h and 24 h), rate of donor adherent and enucleation time of maturate oocyte. The maturation rates of oocyte, fusion rates and cleavation rates of cloned embryos were used to assess the efficiency of different procedures. The maturation rates of ovaries with 3 h exposure was higher than that of 3 to 5 h (60.18% vs 52.50%) (P<0.05). Embryonic development competence had no significant difference (P>0.05). The maturation rates were significantly different between group18 h and 24 h (53.81% vs 89.06%, P<0.01). Embryonic development competence had no significant difference (P>0.05); fusion rates of donor adherent 30% group was higher than that of 10% group. Embryonic development competence had no significant difference (P>0.05). Different adherent donor characterizes the difference in plateau phase. The cleavation rates of 18 hpm group was higher than that of 16 hpm group. Embryonic development competence had no significant difference (P>0.05), the enucleation of 16 hpm group obtained one clone fetus, we got four clone fetus to repeat the 16 hpm group. Five microsatellite was analyzed by PAGE, the bands indicated that fingerprint of cloned fetus were completely the same as those of donor cells. Our data therefore suggests program optimization before enucleation assurance quality of material which be able to improve the quantity and quality of clone embryos, and optimized scheme can obtain clone sheep offspring.

Pedicle screw fixation for scoliosis using Iso-C arm three-dimensional navigation：problems and advantages / 中国组织工程研究

BACKGROUND:Due to variation of spinal anatomic structure, vertebral body rotation, absence of vertebral body, and spinal canal deformation in patients with scoliosis, the deviation of pedicle screw placement can induce catastrophic results. Recently, with the wide application of pedicle screw and the continuous development of computer navigation, scoliosis surgery becomes more accurate and safe. OBJECTIVE:To review the existing problems and advantages of Iso-C arm three-dimensional (3D) navigation system for guiding internal fixation of pedicle screw implantation in the treatment of scoliosis. METHODS: The first author searched PubMed (http://www.ncbi.nlm.nih.gov/PubMed) and CNKI (http://www.cnki.net/) for articles published until December 2012. The key words were “computer navigation, scoliosis, pedicle screw”. The literatures with abstracts containing above key words were retrieved. Totaly more than 50 articles were found, and 13 articles were accorded with the inclusion criteria. RESULTS AND CONCLUSION: Iso-C arm 3D navigation system for guiding internal fixation of pedicle screw implantation in the treatment of scoliosis had obvious advantages. It fuly combined the advantages of CT 3D navigation and intraoperative X-ray navigation, and can quickly and accurately capture the image data. Real time display of images could ensure the safety of repair operation. Automatic registration function of Iso-C arm 3D navigation undoubtedly improves the efficiency and accuracy of the repair, and helps physicians with more accurate and safe ways to carry out a variety of complex scoliosis surgery.

Effects of medicated serum containingDuhuojisheng Decoction on the expression of cytochrome C, proCaspase-9 and proCaspase-3 in degenerative chondrocytes / 中国组织工程研究

BACKGROUND:Previous studies have shown that apoptosis, a central feature of articular chondrocytes, plays a dominant role in cartilage damage, which is one of the pathological factors of articular cartilage degeneration. OBJECTIVE:To observe the effects of meditcated serum containingDuhuojisheng Decoction on the expression of cytochrome C, proCaspase-9 and proCaspase-3 in rat degenerative chondrocytes in vitro and to investigate the possible molecular biological mechanism ofDuhuojisheng Decoction in the treatment of knee osteoarthritis. METHODS:A cultivation system of degenerative chondrocytes in vitro was established. After treatment with meditcated serum containingDuhuojisheng Decoction or blank serum for 24 and 48 hours, the protein expression of cytochrome C, proCaspase-9 and proCaspase-3 was measured by western blot assay. RESULTS AND CONCLUSION: In the cytoplasm, the release of cytochrome C was reduced gradualy in both groups in a time-dependent manner, and the release amount of cytochrome C was significantly lower in the medicated serum group than the blank serum group (P < 0.05). In mitochondria, cytochrome C leakage was gradualy decreased in both groups, and it was decreased significantly in the medicated serum group compared with the blank serum group (P < 0.05). The protein expression of proCaspase-9 and proCaspase-3 was gradualy increased in both groups, especialy in the medicated serum group; the medicated serum containingDuhuojisheng Decoction could promote the protein expression of proCaspase-9 and proCaspase-3 in a time-dependent manner, and there was a significant difference at 24 and 48 hours (P< 0.01). These findings indicate that the medicated serum containingDuhuojisheng Decoction can inhibit the apoptosis of osteoarthritis chondrocytes through inhibiting the release of cytochrome C and the activation of Caspase-9 and Caspase-3.

Direct activation of expression vector withβ-casein gene promoter using engineered TALE transcription factors in mouse fibroblasts / 中国实验动物学报

Objective TALE-TFs were adopted to provide a new way in detection of the expression result ofβ-ca-sein gene promoter-interesting gene expression cassettes in mouse fibroblasts.Methods TALE-TFs of eukaryotic expres-sion plasmid and expression cassette withβ-casein gene promoter and red fluorescent protein reporter gene were co-nucleo-fected into mouse fibroblasts by Amaxa nucleofector.Results and Conclusion β-casein gene promoter was activated by artificial TALE-TFs in the mouse fibroblasts.The way is a new expression verification system instead of mammary epithelial cells with fibroblasts.

Detection and clinical significance of C reaction protein in patients with ulcerative colitis / 中国基层医药

Objective To study the detection and clinical significance of C reaction protein(CRP)in patients with ulcerative colitis(UC).Methods The levels of peripheral blood CRP and blood platelets count(BPC)in patients with UC(n=57) and normal controls group(n=30)were detected and the effects on disease severity were analyzed subsequently.Results The levels of peripheral blood CRP and BPC in active phase group were significantly higher than those in catabasis group(P＜0.05),they were no significant difference between catabasis group and norreal controls group(P＞0.05);the leveh of peripheral blood CRP and BPC in severe stage were significantly higher than those in medium stage(P＜0.05),in medium stage,severe stage they were significantly higher than those in mild stage(P＜0.05,P＜0.01).Conclusion The level of peripheral blood CRP and BPC had important value to judge active phase and severity of UC.

In vitro effect of celecoxib on osteoblast apoptosis induced by titanium particle in neonatal rats / 中国矫形外科杂志

[Objective] To study the effect of celecoxib on osteoblast apoptosis induced by titanium particle in neonatal rats. [Methods]The best in vitro celecoxib serum concentration for promoting proliferation of osteoblast in neonatal rats was obtained through MTT.One group of rat osteoblast in vitro was cultured with serum containing saline and titanium particle and the other was cultured with serum containing celecoxib and titanium particles.Concentrations of TNF-? and PGE-2 in cells collected after 24 hours were analyzed by ELLSIA,COX-2 mRNA and protein were indicated by FO-PCR and Westren blot.[Results]Some cellar factors such as COX-2,TNF-? and PGE-2 could be induced by titanium particle in in vitro,but celecoxib could inhibit expression of these factors by COX-2 pathway.[Conclusion]Celecoxib coule be a promising drug for treatment of aseptic loosening.

Treating Hp-related digestive ulcer with esomeprazole / 中国基层医药

Objective To investigate the effect of esomeprazole in the treatment of digestive ulcer with heli- eobacter pylori(Hp)infection.Methods A total of 60 patients with digestive ulcer and Hp infection were randomly divided into group A(30 cases)treated with esomeprazole(20mg),amoxicillin(1000mg)and clarithromycin(500 mg)twice daily and group B(30 cases)treated with omeprazole(20mg),amoxicillin(1000mg)and clari thromycin (500mg)twice daily.The treatment lasted for 7 days,after which abdominal pain refief rate,ulcer healing rate and eradication rate of Hp as well as adverse effects of the medication were evaluated and compared.Results Ulcer heal- ing rate and Hp eradication rate were 90.1% and 94,2 % in group A and 83.9 % and 83.7 % in group B,showing significant statistical diferences between the two groups(P