Genetic modification technology is a new molecular tool for targeted genome modification. It includes zinc finger nucleases (ZFN) technology, transcription activator-like effector nucleases (TALEN) technology and clustered regularly interspaced short palindromic repeat (CRISPR)-associated (Cas) (CRISPR-Cas) nucleases technology. All of these nucleases create DNA double-strand breaks (DSB) at chromosomal targeted sites and induce cell endogenous mechanisms that are primarily repaired by the non-homologous end joining (NHEJ) or homologous recombination (HR) pathway, resulting in targeted endogenous gene knock-out or exogenous gene insertion. In recent years, genetic modification technologies have been successfully applied to bacteria, yeast, human cells, fruit fly, zebra fish, mouse, rat, livestock, cynomolgus monkey, Arabidopsis, rice, tobacco, maize, sorghum, wheat, barley and other organisms, showing its enormous advantage in gene editing field. Especially, the newly developed CRISPR-Cas9 system arose more attention because of its low cost, high effectiveness, simplicity and easiness. We reviewed the principles and the latest research progress of these three technologies, as well as prospect of future research and applications.
Animals ; CRISPR-Cas Systems ; DNA Breaks, Double-Stranded ; Endonucleases ; Genetic Engineering ; methods ; Humans ; Mutagenesis, Insertional ; Mutagenesis, Site-Directed ; Plants ; Zinc Fingers

In recent years, ultrasound elastography, as a new technique for evaluating soft tissue elasticity, has been progressively used in musculoskeletal system. Shear-wave elastography (SWE) is considered to be more objective, quantitative, and reproducible than other ultrasonic elastography techniques with increasing applications to the musculoskeletal system. A number of studies have shown that SWE has high application value in determining severity and prognosis of the musculoskeletal tissue diseases (including tendons, muscles, nerves and ligaments). This article describes the applications of SWE in the evaluation of musculoskeletal system.

PURPOSE: MicroRNAs are small non-coding RNAs that play important roles in vascular smooth muscle cell (VSMC) function. This study investigated the role of miR-379 on proliferation, invasion, and migration of VSMCs and explored underlying mechanisms thereof. MATERIALS AND METHODS: MicroRNA, mRNA, and protein levels were determined by quantitative real-time PCR and western blot. The proliferative, invasive, and migratory abilities of VSMCs were measured by CCK-8, invasion, and wound healing assay, respectively. Luciferase reporter assay was used to confirm the target of miR-379. RESULTS: Platelet-derived growth factor-bb was found to promote cell proliferation and suppress miR-379 expression in VSMCs. Functional assays demonstrated that miR-379 inhibited cell proliferation, cell invasion, and migration. Flow cytometry results further showed that miR-379 induced apoptosis in VSMCs. TargetScan analysis and luciferase report assay confirmed that insulin-like growth factor-1 (IGF-1) 3'UTR is a direct target of miR-379, and mRNA and protein levels of miR-379 and IGF-1 were inversely correlated. Rescue experiments showed that enforced expression of IGF-1 sufficiently overcomes the inhibitory effect of miR-379 on cell proliferation, invasion, and migration in VSMCs. CONCLUSION: Our results suggest that miR-379 plays an important role in regulating VSMCs proliferation, invasion, and migration by targeting IGF-1.
Apoptosis ; Cell Movement/*physiology ; Cell Proliferation/*physiology ; Humans ; Insulin ; Insulin-Like Growth Factor I/*physiology ; MicroRNAs/*physiology ; Muscle, Smooth, Vascular/*cytology ; Proto-Oncogene Proteins c-sis/*physiology ; RNA, Messenger/metabolism ; Real-Time Polymerase Chain Reaction ; Sincalide/physiology ; Wound Healing/physiology

Objective:To examine the role of LY294002 in cardiac function and myocardial structure in dilated cardiomyopathy (DCM) rats.Methods:Fifty-two male SD rats were randomly assigned to a control group (n=16) and a DCM group (n=36).The DCM rats were induced by intraperitoneal injection of adriamycin,and the control rats were given normal saline.After observation for 2 weeks,6 rats from each group were killed randomly.In the end of the 8th week,the 24 DCM rats were randomly assigned to a DCM group (n=12) and a LY294002 group (n=12),which were given normal saline and LY294002,respectively.In the end of the 8th week and 16th week,the cardiac function was analyzed by ultrasonic cardiogram (UCG) and the plasma was collected to test the level of N-terminal pro-brain natriuretic peptide (NT-pro BNP).HE and Van Gieson (VG) staining were performed to calculate the collagen volume fraction (CVF).Results:Compared with the control group,the left ventricular end-diastolic dimension (LVEDD),left ventricular end-systolic dimension (LVESD) and NT-proBNP level of in the DCM rats were increased obviously,while the left ventricular ejection fraction (LVEF),left ventricular fractional shortening (LVFS) in the DCM rats were decreased obviously (P＜0.01).These changes were consistent with DCM characteristics.Compared with the DCM group,the LVEDD,LVESD and NT-proBNP levels in the LY294002 group were decreased,while the LVEF and LVFS were increased (P＜0.05).Histopathology showed that the myocardium in the DCM rats was fibrotic and the CVF was increased compared with the control rats (P＜0.01).The myocardial structure was improved in the LY294002 group compared to the DCM group.Conclusion:LY294002 can reduce the myocardial fibrosis in the DCM rats and improve the cardiac function.

Objective To observe the changes of cell cycle,apoptosis and proliferation of endometrial cancer cells after the expression and down-regulation of Ezrin in endometrial cancer cells and to explore whether Ezrin may be a candidate gene for targeted therapy.Methods Endometrial cancer cells were from Shanghai Institute of Cell Research,of Chinese Academy of Medical Sciences in February 2017 and divided into blank control group and siEzrin group according to the intervention methods.Western blot and qRT-PCR was used to detect the expression of Ezrin protein and mRNA in endometrial cell lines.Small interfering RNA (siRNA) was used to transfect HEC-1B cell and down-regulate Ezrin.Cell cycle and apoptosis were detected by flow cytometry.MTT assay was used to detect multiplication.Results Western blot showed that Ezrin protein was expressed in Ishikawa (31.742 ± 5.832)、HEC-1A (16.326 ± 3.135)、HEC-1B(17.636±4.426) and KLE(14.862±5.109) and qRT-PCR showed that mRNA was expressed in Ishikawa (2.513±0.725),HEC-1A (1.655±0.692),HEC-1B (3.237±0.411) and KLE (0.962±0.235) cell lines,and expressed highest in HEC-1B cells (F=6.173,P＜0.05;F=7.042,P＜0.05).Flow cytometry assay showed that compared with blank control group less cells stayed in G1 phase and G2 phase,more stayed in S phase (t=3.118,P＜0.05;t=5.435,P＜0.05;t=3.332,P＜0.05).The apoptotic rate of HEC-1B cells increased from (9.84 ± 2.37) ％ to (17.64 ± 5.96) ％ (t =8.963,P ＜ 0.01) after Ezrin was downregulated.MTT assay showed that the proliferation of HEC-1B cells in 72 h and 96 h siEzrin transfection group was lower than that in blank control group (t =3.209,P＜ 0.05;t =3.726,P＜ 0.05).Conclusion Down-regulating of Ezrin may promote more endometrial cancer cells stay in S phase and promote apoptosis,inhibit proliferation,Ezrin may become target candidate gene in target therapy.

The role of malate dehydrogenase 2 (MDH2) in tumors is double-sided,it has a cancerpromoting effect in some tumors and an inhibitory effect in other tumors.The function of MDH2 is related to energy metabolism,tumor resistance and its pseudo hypoxia.MDH2 plays an important role in the occurrence,development,invasion and metastasis of tumors.An in-depth understanding of the functional mechanism of MDH2 in tumors can provide new molecular targets for tumor intervention in the clinic.

To evaluate the volume and function of left atrium and left atrial appendage in patients with atrial fibrillation by three-dimensional transesophageal echocardiography and transthoracic echocardiography.
 Methods: A total of 112 patients with atrial fibrillation were divided into two groups: a paroxysmal atrial fibrillation (ParAF) group (n=80) and a persistent atrial fibrillation (PerAF) group (n=32). Control group was people without atrial fibrillation (n=40). Clinical data of the participants were collected. Left atrial dimension (LAD), left atrial volume (LAV), left ventricular end-diastolic volume (LVEDV) and left ventricular end-systolic volume (LVESV) were measured by transthoracic echocardiography, while left atrial appendage peak emptying flow velocity (LAAeV), left atrial appendage peak filling flow velocity (LAAfV), left atrial appendage maximum volume (LAAVmax) and left atrial appendage minimum volume (LAAVmin) were measured by three-dimensional transesophageal echocardiography. Left atrial volume index (LAVI), left ventricular ejection fraction (LVEF) and left atrial appendage ejection fraction (LAAEF) were calculated. 
 Results: Compared with the control group, LAAEF, LAAeV and LAAfV in the ParAF group were decreased obviously, while LAD, LAV, LAVI, LAAVmax and LAAVmin in the ParAF group were increased obviously (P<0.05). Compared with the ParAF group, LAAEF, LAAeV and LAAfV in the PerAF group were also decreased obviously, and LAD, LAV, LAVI, LAAVmax and LAAVmin in the ParAF group were also increased obviously (P<0.05). There was no statistically significant difference in LVEDV, LVESV, LVEF between the ParAF group and the PerAF group (P<0.05).
 Conclusion: Left atrium and left atrial appendage were enlarged and the function of left atrial appendage was declined in patients with AF, and the changes were more obvious in patients with PerAF compared with patients with ParAF by three-dimensional transesophageal echocardiography and transthoracic echocardiography.
Atrial Appendage ; physiopathology ; Atrial Fibrillation ; physiopathology ; Echocardiography, Transesophageal ; Heart Atria ; physiopathology ; Humans

Hypersplenism is an important complication of cirrhotic portal hypertension, and splenectomy is an important means to treat hypersplenism in cirrhosis. It is realized that hypersplenism played a pathological role in the course of cirrhosis. This article analyzes and compares the changes in the condition of patients with cirrhosis between splenectomy with and without hyperfunction, and comprehensively discusses the pathological role and mechanism of hypersplenism in the course of cirrhosis, in order to strengthen the clinical prevention and treatment of hypersplenism in cirrhosis and to better improve the condition and prognosis of patients with cirrhosis.

Objective To investigate the frailty status and influencing factors of the elderly stroke pa-tients in community to provide a basis for its prevention and treatment.Methods The convenience sampling was used to select 200 elderly stroke patients as the study subjects in a community hospital in Guangzhou City from October 2020 to January 2022.The general information questionnaire,Chinese version of Tilburg frailty indicator scale,Modified Rankin Index Scale,Barthel index scale,short version of Geriatric Depression Scale and Social Support Rating Scale were used to collect the data.Results The incidence rate of frailty in 200 ca-ses of community elderly stroke was 86.0％.The results of multiple linear regression analysis showed that the degree of disability,ability of daily living,depression and social support were the influencing factors of frailty of the elderly stroke patients in community(F=71.813,P＜0.001).Conclusion The incidence rate of frailty is higher in community elderly stroke patients,the higher the degree of disability,the lower the ability of daily living,the higher the level of depression and the lower the level of social support,the higher the frailty level of community elderly stroke patients.

AIM: To explore the value of the corneal/refractive analyzer OPD-Scan III in assessing visual quality after Nd:YAG laser surgery in patients with posterior capsular opacification.METHODS: A total of 98 patients(98 eyes)with posterior capsular opacification who underwent Nd:YAG laser posterior capsulotomy in our hospital from May 2021 to April 2023 were retrospectively selected as the research subjects, and higher-order aberration parameters(coma, spherical aberration and trefoil aberration)and visual quality parameters [Strehl ratio(SR)and 4 mm area ratio(AR)in the human eye modulation transfer function(MTF)curve] were obtained using the corneal/refractive analyzer OPD-Scan III. The patient's visual acuity, higher-order aberration parameters and visual quality before surgery, and at 1 wk and 3 mo after surgery were compared.RESULTS: There were statistically significant differences in uncorrected distance visual acuity(UDVA), uncorrected intermediate visual acuity(UIVA)and uncorrected near visual acuity(UNVA), coma, spherical aberration, trefoil aberration, SR and AR before and after surgery(all P<0.001), and there were statistically significant differences in UDVA, UIVA, UNVA, coma, spherical aberration, trefoil aberration, SR and AR at 1 wk and 3 mo postoperatively(all P<0.05). Additionally, there were differences in the UDVA, UIVA, UNVA, SR and AR at 3 mo postoperatively compared with those parameters at 1 wk postoperatively(all P<0.05), while there were no statistical significance in coma, spherical aberration, trefoil aberration at 3 mo postoperatively and those at 1 wk postoperatively(all P>0.05). Furthermore, the spherical aberration, trefoil aberration were negatively correlated with SR(rs=-0.427, P=0.009; rs=-0.436, P=0.010), and the trefoil aberration was negatively correlated with AR(rs=-0.406, P=0.015).CONCLUSION: The visual acuity of patients with posterior capsular opacification improved after Nd:YAG laser surgery, and the corneal/refractive analyzer OPD-Scan III system has a high clinical value in reflecting patient's visual quality.