Mammals exhibit limited heart regeneration ability, which can lead to heart failure after myocardial infarction. In contrast, zebrafish exhibit remarkable cardiac regeneration capacity. Several cell types and signaling pathways have been reported to participate in this process. However, a comprehensive analysis of how different cells and signals interact and coordinate to regulate cardiac regeneration is unavailable. We collected major cardiac cell types from zebrafish and performed high-precision single-cell transcriptome analyses during both development and post-injury regeneration. We revealed the cellular heterogeneity as well as the molecular progress of cardiomyocytes during these processes, and identified a subtype of atrial cardiomyocyte exhibiting a stem-like state which may transdifferentiate into ventricular cardiomyocytes during regeneration. Furthermore, we identified a regeneration-induced cell (RIC) population in the epicardium-derived cells (EPDC), and demonstrated Angiopoietin 4 (Angpt4) as a specific regulator of heart regeneration. angpt4 expression is specifically and transiently activated in RIC, which initiates a signaling cascade from EPDC to endocardium through the Tie2-MAPK pathway, and further induces activation of cathepsin K in cardiomyocytes through RA signaling. Loss of angpt4 leads to defects in scar tissue resolution and cardiomyocyte proliferation, while overexpression of angpt4 accelerates regeneration. Furthermore, we found that ANGPT4 could enhance proliferation of neonatal rat cardiomyocytes, and promote cardiac repair in mice after myocardial infarction, indicating that the function of Angpt4 is conserved in mammals. Our study provides a mechanistic understanding of heart regeneration at single-cell precision, identifies Angpt4 as a key regulator of cardiomyocyte proliferation and regeneration, and offers a novel therapeutic target for improved recovery after human heart injuries.
Humans ; Mice ; Rats ; Cell Proliferation ; Heart/physiology* ; Mammals ; Myocardial Infarction/metabolism* ; Myocytes, Cardiac/metabolism* ; Pericardium/metabolism* ; Single-Cell Analysis ; Zebrafish/metabolism*

As main recipient cells for porcine reproductive and respiratory syndrome virus (PRRSV), porcine alveolar macrophage (PAM) are involved in the progress of several highly pathogenic virus infections. However, due to the fact that the PAM cells can only be obtained from primary tissues, research on PAM-based virus-host interactions remains challenging. The improvement of induced pluripotent stem cells (iPSCs) technology provides a new strategy to develop IPSCs-derived PAM cells. Since the CD163 is a macrophage-specific marker and a validated receptor essential for PRRSV infection, generation of stable porcine induced pluripotent stem cells lines containing CD163 reporter system play important roles in the investigation of IPSCs-PAM transition and PAM-based virus-host interaction. Based on the CRISPR/Cas9- mediated gene editing system, we designed a sgRNA targeting CD163 locus and constructed the corresponding donor vectors. To test whether this reporter system has the expected function, the reporter system was introduced into primary PAM cells to detect the expression of RFP. To validate the low effect on stem cell pluripotency, we generated porcine iPSC lines containing CD163 reporter and assessed the pluripotency through multiple assays such as alkaline phosphatase staining, immunofluorescent staining, and EdU staining. The red-fluorescent protein (RFP) expression was detected in CD163-edited PAM cells, suggesting that our reporter system indeed has the ability to reflect the expression of gene CD163. Compared with wild-type (WT) iPSCs, the CD163 reporter-iPSCs display similar pluripotency-associated transcription factors expression. Besides, cells with the reporter system showed consistent cell morphology and proliferation ability as compared to WT iPSCs, indicating that the edited-cells have no effect on stem cell pluripotency. In conclusion, we generated porcine iPSCs that contain a CD163 reporter system. Our results demonstrated that this reporter system was functional and safe. This study provides a platform to investigate the iPS-PAM development and virus-host interaction in PAM cells.
Swine ; Animals ; Induced Pluripotent Stem Cells/metabolism* ; Receptors, Cell Surface/genetics* ; Antigens, CD/metabolism* ; Porcine respiratory and reproductive syndrome virus/genetics*

Objective To investigate the efficacy of acupuncture and rehabilitation therapy on lower limb motor function, and to explore a cortical mechanism using functional near infrared spectroscopy (fNIRS). Methods From December, 2020 to July, 2021, 24 stroke patients with lower limb motor dysfunction in our hospital were randomly divided into rehabilitation group (n = 12) and acupuncture-rehabilitation group (n = 12), and received routine rehabilitation training and acupuncture-rehabilitation intervention for four weeks, respectively. The control group included ten healthy subjects matched the patients. Before and after intervention, the lower limb motor function of the patients was assessed with Fugl-Meyer Assessment-Lower Extremities (FMA-LE), and all the subjects accepted fNIRS examination. The functional intensity and lateralization index (LI) of supplementary motor area (SMA), premotor cortex (PMC) and sensory motor cortex (SMC) were calculated based on oxygenated hemoglobin (HbO2). Results There was no significant difference in FMA-LE score between the rehabilitation group and the acupuncture-rehabilitation group before the intervention (P > 0.05). After four weeks of intervention, FMA-LE scores improved in both groups (t > 3.770, P < 0.001), and improved more in the acupuncture-rehabilitation group than in the rehabilitation group (t = 2.252, P < 0.05). Before intervention, the average functional connection was more intensitive in the control group than in the two patients groups (P < 0.05), and there was no significant difference between the later two groups (t = 0.458, P > 0.05). After intervention, the average functional connection increased in both groups (t > 2.178, P < 0.05), and the functional connection of the affected PMC of acupuncture-rehabilitation group increased (P < 0.05). The LI in SMC increased in the acupuncture-rehabilitation group (P < 0.05). There was a significant positive correlation between the change of functional connection of the affected PMC and the change of FMA-LE scores in the acupuncture-rehabilitation group (r = 0.579, P < 0.05). Conclusion Acupuncture with rehabilitation therapy can significantly improve the lower limb motor function and asymmetrical activation of SMC in stroke patients. The recovery of lower limb motor function may be related to the enhanced activation of affected PMC.

TORCH, which is considered as a series of pathogens, including the Toxoplasma gondii, Rubella virus, Cytomegalovirus or Herpes simplex virus, often infects the pregnant women to induce the the fetus or newborn infection by transplacental infection or exposure to contaminated genital tract secretions at delivery. Increasing evidence have been confirmed that the infection of TORCH may cause the miscarriage, premature birth, malformed fetus, stillbirth, intrauterine growth retardation, neonatal multiple organ dysfunction and other adverse pregnancy outcomes. For most TORCH-infections cases may lacking the effective treatments during pregnancy, and it is important to achieve the effacing monitoring of TORCH infections before and during pregnancy. The laboratory testing of TORCH has the great significance. However, the consensus opinions still need to improve the the standardization of TORCH testing process and the correct interpretation. Based on the characteristics of the TORCH detection method, this article gives a consensus opinion on the standardized detection and clinical application of TORCH from the laboratory perspective according to the characteristics and types of infection of different pathogens.

Objective    To evaluate the prognosis of interventional treatment with covered stent graft for retrograde Stanford type A aortic dissection and intramural hematoma by single-arm meta-analysis. Methods    Related studies on treating retrograde Stanford type A aortic dissection and intramural hematoma with covered stent graft were retrieved from the databases by computer, including PubMed, EMbase, The Cochrane Library, Wanfang Data, VIP, CNKI and CBM, from inception to January 2020. Literatures were screened by researchers step by step according to the predefined inclusion and exclusion criteria. Quality of the enrolled literatures was evaluated, and data were extracted from the included studies. Afterwards, single-arm meta-analysis was carried out by the R3.6.3 software. Results    A total of 12 English and 5 Chinese studies were included, which were all case series, and the quality of all literatures was moderate evaluated by Newcastle-Ottawa Scale (NOS). After analyzing the clinical prognosis of 260 patients, the 30-day mortality was 6% (95%CI 0.04 to 0.11, P=0.97), the late mortality was 8% (95%CI 0.05 to 0.14, P=0.78), the incidence of endoleak was 21% (95%CI 0.16 to 0.29, P=0.06), the incidence of stroke was 5% (95%CI 0.03 to 0.09, P=0.99), the incidence of new aortic dissection was 7% (95%CI 0.04 to 0.11, P=0.96), the incidence of dissection progression was 10% (95%CI 0.07 to 0.16, P=0.24), and the absorption rate of intramural hematoma was 84% (95%CI 0.37 to 1.00, P<0.01). Conclusion    Interventional treatment with covered stent graft for retrograde Stanford type A aortic dissection and intramural hematoma can obtain good early treatment results for some patients, and can be used as a safe and effective treatment for aged patient with high risk who cannot tolerate surgery. Endoleak, stroke and new aortic dissection are the early serious complications of this method.

Objective To study the thrombolysis effect and safety of UK combined with tirofiban in young patients with ST segement elevation acute myocardial infarction.Methods 76 young patients with ST segement elva-tion acute myocardial infarction were selected as research objects,and they were divided into UK group(control group, n =34)and UK combined with tirofiban group(observation group,n =42)by random number table,then the recanali-zation rate of infarct -related artery and incidence of complications were compared.Results The recanalization rate of infarct -related artery in the observation group was 97.6%,which was 91.2% in the control group,there was no significant difference between the two groups(χ2 =1.564,P >0.05);The vascular infarction related to blood flowⅡlevel of the patients was 90.5% in the observation group,which was significantly higher than 70.6% in the control group(χ2 =4.945,P <0.05);The death,complications incidences had not significant differences between the two groups(χ2 =1.252,2.837,2.837,all P >0.05).Conclusion The thrombolysis effect of UK has a high recanaliza-tion rate of infarct -related artery in young patients with ST segement elevation acute myocardial infarction,and the thrombolysis effect of UK combined with tirofiban is better,and it does not increase the incidence of complications.

Objective To evaluate the incidence and significance of perineural invasion (PNI) in cervical cancer. Methods Retrospective chart review of patients with cervical cancer (stages Ⅰa2-Ⅱb) who underwent radical hysterectomy and pelvic lymphadenectomy from 2007 to 2012. To evaluate the incidence and significance of PNI in cervical and uterine tissues by microscopic examination. Results A total of 238 patients were included, 9.2% (22/238) patients with PNI in the cervical stroma. Patients with PNI were more likely to have adverse histopathologic features, including lymphoma vascular space invasion, parametrical invasion, depth of invasion, tomor size and lymph nodes metastases (all P<0.05). PNI were independent of age, international federation of gynecology and obstetrics (FIGO) stage, histopathology type and grade, and positive vaginal margin (all P>0.05). Patients with PNI had shorter disease-free and overall survival (P=0.002 and P=0.008, respectively). On multivariate analysis, risk factors for recurrence and death included parametrical invasion and depth of invasion (P<0.05). Similarly, risk factors for recurrence included lymph nodes metastases (P=0.024). However, PNI was not identified as an independent risk factor for either recurrence or death (P>0.05). Conclusions PNI exists in early cervical cancer. PNI is associated with tumor size, depth of invasion, parametrical invasion, lymphoma vascular space invasion and lymph nodes metastases. PNI represente a decreasing disease-free and overall survival in patients with early-stage cervical cancer, and is independently associated with multiple high-risk factors, which be informed management decisions regarding adjuvant therapy.

Objective To evaluate the value of the application of ultrasonic elastography and ultrasound contrast imaging on thyroid microcarcinoma(TMC). Methods The in-patients with thyroid lesions admitted in the Second People's Hospital of Foshan City from January 2013 to April 2014 were enrolled. Two-dimensional ultrasonography was used to screen the 162 solid thyroid nodules with a diameter ≤10 mm;they were examined by ultrasonic elastography and ultrasound contrast imaging respectively,and the result of pathological examination was regarded as the golden standard to compare the sensitivity and specificity between ultrasonic elastography and ultrasound contrast imaging for the diagnosis of TMC. Results The pathological results showed there were benign nodules 118 and malignant nodules 44 in number. The sensitivity,specificity,incidence of accuracy,positive predictive value and negative predictive value of the diagnosis of TMC by ultrasonic elasticity imaging,ultrasound imaging,and elasticity imaging combined with ultrasound imaging were much higher than those by two-dimensional ultrasonography〔sensitivity:88.64%(39/44),90.91%(40/44),95.45%(42/44)vs. 81.82%(36/44);specificity:91.53%(108/118),92.37%(109/118),95.76%(113/118)vs. 85.59%(101/118);accuracy:90.74%(147/162), 91.97%(149/162),93.26%(151/162)vs. 84.56(137/162);positive predictive value:79.59%(39/49),81.63%(40/49),83.67%(41/49)vs. 67.92%(36/53);negative predictive value:95.58%(108/113),96.46%(109/113), 97.35%(110/113)vs. 92.66%(101/109),all P<0.05〕. Conclusion For the diagnosis of TMC,both ultrasonic elastography and ultrasound contrast imaging have rather high value,and when they are combined together to be applied for the diagnosis,the result is the best.

Objective To characterize the feasibility of low-dose CT angingraphy on 16-slice multislice computed tomography (16-MSCT), and its relationship to the noise in the pre-contrast image and enhancement value.Methods Forty-three consecutive patients (male 21, female 22, mean age 59 years, median age 56 years) underwent abdominal 16-MSCT (Toshiba Aquilion 16) with constant scanning parameters including 120 kVp, a 0.5-second gantry rotation time, a pitch of 0.938: 1, and 16×1-mm detector collimation.The mA was set at 200 in the pre-contrast scan and 160 in the contrast-enhanced scan.The arterial phase images were retrospectively reconstructed with 1-mm slice thickness, 0.8 mm interval.The pre-contrast noise was defined as the standard deviation (SD) of the aorta at the level of right posterior crura of diaphragm.The enhancement of aorta was also measured at level of celiac artery.The volume rendering of CT angiography was made and classified into three grades (excellent, good, bad).Receiver operating characteristic curve (ROC) was used to evaluate the relationship between the image quality of CT angiography and noise in the pre-contrast image and enhancement value.Results Twenty-five cases had the aorta enhancement between 300.0--400.0 HU.The sensitivity and specificity of excellent CTA image was 75% and 62%, respectively when the SD was 12.00.Eighteen cases had the aorta enhancement more than 400.0 HU.The sensitivity and specificity of excellent CTA image was all 100% when the SD was 12.25, and 100% and 75%, respectively when the SD was 13.35.The area under curve of CTA image quality and enhancement in receiver-operated characteristic analysis was O.907.The enhancement was more than 356.7 HU when the sensitivity and specificity of excellent CTA image was 100% and 60%, respectively, and 389.8 HU when 78% and 80%.When the enhancement was more than 442.4 HU, the specificity of excellent CTA image was 100%.Conclusions MSCT angiography with low radiation dose is feasible.Good CT angiography demands great enhancement, and then, the needed mAs is lower when the SD is smaller.

Objective To establish a method for rapid diagnosis and identification of seven species human herpesviruses infection.Methods Primers and oligonucleotide probes were designed and synthesized based on the highly conserved regions of the DNA polymerase genes in human herpesviruses, namely herpes simplex virus type 1 (HSV-1),HSV-2,varicella-zoster virus (VZV),Epstein-Barr virus (EBV),cytomegalovirus (CMV),and human herpes virus 6 (HHV-6A/6B).DNA microarrays were made by printing the oligonucleotide probes on the special glass slides.A total of 282 blood specimens from children with suspected infection were analyzed by this DNA microarray technique,and the results were compared with those of TaqMan PCR.Results The products of the seven human herpesviruses after PCR amplification could be used to identify the virus species with DNA microarrays.The detection limits were 10 copies/?l for HSV-1,HSV-2,VZV,EBV,CMV,HHV-6A,and HHV-6B,respectively.The assay did not show cross-reaction to the DNA extract of hepatitis B virus,staphylococcus aureus,E.coli,Candia albicans and human genome.Among the 282 samples,59 were positive for human herpesviruses DNA.Compared with those of TaqMan PCR,the sensitivity and specificity of the microarray assay were 96.7% and 99.5%,respectively,and the index of accurate diagnosis was 0.962.Conclusions This DNA microarray for identifying human herpesviruses species is specific and sensitive,and may serve as an efficient technique for simultaneous detection and species identification of human herpesviruses in clinical specimen.