Chimeric antigen receptor (CAR) T-cell therapy has achieved breakthroughs in treating relapsed/refractory B-cell malignancies. However, it still faces challenges, including complex manufacturing processes, limited indications, T-cell exhaustion, and insufficient durability of therapeutic efficacy. CRISPR/Cas9, a highly efficient and relatively simple gene-editing technology, offers new avenues for overcoming these limitations. This review briefly outlines the working mechanism of CRISPR/Cas9 and focuses on its recent applications and clinical practices in developing universal CAR T-cells, enhancing T-cell function, and extending CAR T-cell therapy to T-cell and myeloid leukemias. Furthermore, this review highlights optimization strategies developed over the past two years to enhance the editing precision, delivery efficiency, and safety of the CRISPR/Cas9 system, aiming to provide insights for the optimal design and clinical application of CAR T-cell therapy.

Chimeric antigen receptor (CAR) T-cell therapy has achieved breakthroughs in treating relapsed/refractory B-cell malignancies. However, it still faces challenges, including complex manufacturing processes, limited indications, T-cell exhaustion, and insufficient durability of therapeutic efficacy. CRISPR/Cas9, a highly efficient and relatively simple gene-editing technology, offers new avenues for overcoming these limitations. This review briefly outlines the working mechanism of CRISPR/Cas9 and focuses on its recent applications and clinical practices in developing universal CAR T-cells, enhancing T-cell function, and extending CAR T-cell therapy to T-cell and myeloid leukemias. Furthermore, this review highlights optimization strategies developed over the past two years to enhance the editing precision, delivery efficiency, and safety of the CRISPR/Cas9 system, aiming to provide insights for the optimal design and clinical application of CAR T-cell therapy.

Objective To investigate the effects of microRNA-26a-5p(miR-26a-5p)on high glucose-induced retina Müller activation and apoptosis by regulating PTEN/PI3K/Akt signaling pathway,and the potential mechanism of diabetic retinal neurodegeneration.Methods Various concentrations of high glucose were added into rMC-1 culture.CCK-8 and flow cytometry was used to examine cell proliferation and apoptosis respectively.The regulatory effects of miR-26a-5p on the expressions of PTEN,PI3K and Akt were observed by real-time PCR;the expression levels of IL-1β and IL-6 in Müller cells were examined by ELISA.The data were processed by Graphpad 8.0 software.Results Müller cells grew actively in high-glucose stimulation culture.Compared with the control group,the activity of Müller cells stimulated by 50 mmol/L glucose increased gradually at 12 h and 24 h,but decreased at 48 h after stimulation,when Müller cells apoptosis increased.The difference between the groups was statistically significant(P＜0.05).The expression of miRNA-26a-5p decreased,that of PTEN increased,and those of PI3K and Akt decreased.Meanwhile,IL-1β and IL-6 levels were significantly increased in Müller cells.miR-26a-5p over-expression alleviated injuries to high glucose stimulated retinal Müller cells by inhibiting PTEN,which upregulated the expression of PI3K/Akt and downregulation of IL-1β and IL-6(P＜0.01).Conclusion Upregulating miR-26a-5p protects Müller cells against apoptosis,probably through regulation of PTEN/PI3K/Akt and affecting the production of inflammatory factors.

AIM: To study the protective effect of fenofibrate on diabetic retinal neurodegeneration and observe its effect on miR-26a-5p and its target gene PTEN in the retinal of diabetic mice.METHODS: Diabetic mice models were established and they were gavaged by fenofibrate. H& E staining and transmission electron microscopy were used to observe the impairments of retinal neurons. Real-time PCR was used to examine the expression of miR-26a-5p, and Western blotting was employed to measure the expression of phosphatase and tensin homologue(PTEN)in the retina of diabetic mice. The expression level of nuclear factor-κB(NF-κB), interleukin-1β(IL-1β)and the morphology of neural tissues were observed.RESULTS: When compared with the diabetic mice, fenofibrate significantly attenuated the damage to retinal ganglion cells and the atrophy of retinal nerve fiber layer. While the level of miR-26a-5p was increased and the levels of PTEN and inflammatory mediators were significantly decreased in the retina of fenofibrate treated diabetic mice, with significant statistical significance(P<0.05).CONCLUSIONS: Fenofibrate protects against diabetic retinal neurodegeneration by upregulating miR-26a-5p and inhibiting PTEN, attenuating the inflammatory response and alleviating retinal cell injury.

Background::Genetic variants of dopaminergic transcription factor-encoding genes are suggested to be Parkinson’s disease (PD) risk factors; however, no comprehensive analyses of these genes in patients with PD have been undertaken. Therefore, we aimed to genetically analyze 16 dopaminergic transcription factor genes in Chinese patients with PD.Methods::Whole-exome sequencing (WES) was performed using a Chinese cohort comprising 1917 unrelated patients with familial or sporadic early-onset PD and 1652 controls. Additionally, whole-genome sequencing (WGS) was performed using another Chinese cohort comprising 1962 unrelated patients with sporadic late-onset PD and 1279 controls.Results::We detected 308 rare and 208 rare protein-altering variants in the WES and WGS cohorts, respectively. Gene-based association analyses of rare variants suggested that MSX1 is enriched in sporadic late-onset PD. However, the significance did not pass the Bonferroni correction. Meanwhile, 72 and 1730 common variants were found in the WES and WGS cohorts, respectively. Unfortunately, single-variant logistic association analyses did not identify significant associations between common variants and PD. Conclusions::Variants of 16 typical dopaminergic transcription factors might not be major genetic risk factors for PD in Chinese patients. However, we highlight the complexity of PD and the need for extensive research elucidating its etiology.

Objective To observe the thickness changes in the peripapillary retinal nerve fiber layer(RNFL)and the ganglion cell-inner plexiform layer(GC-IPL)in the macular region,so as to analyze the pathological rules of retinal nerve injury in diabetes retinopathy(DR)patients and explore the related risk factors.Methods A total of 164 patients(164 eyes)with type 2 diabetes who visited the Department of Ophthalmology and Department of Endocrinology of Shaanxi Pro-vincial People's Hospital from January 1,2017 to January 1,2018 were selected as the subjects.According to the condition of concomitant DR,these subjects were divided into three groups:non-DR(NDR)group(56 eyes of 56 patients),mild non-proliferative DR(NPDR)group(53 eyes of 53 patients)and moderate NPDR group(55 eyes of 55 patients).Healthy individuals who underwent physical examinations in the same period were selected as the control group(50 eyes of 50 pa-tients).Optical coherence tomography and optical coherence tomography angiography were used to measure the peripapil-lary RNFL,macular GC-IPL,and macular vessel density(VD)of all subjects.Comparison among groups and correlation analysis were conducted on the above retinal nerve injury indicators.Results Compared with the control group and NDR group,the average,upper and lower peripapillary RNFL in the mild NPDR group and moderate NPDR group were signifi-cantly thinner,and the differences were statistically significant(all P＜0.05).The differences in macular GC-IPL thickness at the average,upper,upper temporal,lower temporal,lower,lower nasal,and upper nasal regions in all groups were sta-tistically significant(all P＜0.05);pairwise comparisons showed that compared with the control group,the GC-IPL thick-ness in each quadrant of the macular area of patients in the NDR group,mild NPDR group and moderate NPDR group de-creased significantly(all P＜0.05),with the upper GC-IPL thinning being the most significant;compared with the NDR group,the GC-IPL thickness in the upper,average,and upper temporal macular area of patients with mild NPDR decreased significantly(all P＜0.05);compared with the mild NPDR group,the GC-IPL thickness in the upper macular area was thin-ner in the moderate NPDR group,and the difference was statistically significant(P＜0.05).There were statistically signifi-cant differences in the average and inner ring VDs in the macular area among the groups(both P＜0.05).The correlation analysis results showed that there was a significantly positive correlation between the peripapillary RNFL thickness and the average VD in the macular area(r=0.517,P＜0.01),while the average GC-IPL thickness in the macular area was weakly positively correlated with the average VD in the macular area(r=0.279,P＜0.01).Conclusion The peripapillary RNFL thickness objectively reflects the degree of retinal neuron injury,and is a key index to evaluate the degree of retinal nerve injury in diabetes patients.

Objective:To analyze the curative effect of locking compression pediatric hip plate (LCH-PHP) in the treatment tumor-like lesions of femoral neck in children.Methods:From January 2012 to June 2019, 45 children with Erol type Ⅱ-Ⅳ tumor-like lesions of femoral neck were retrospectively analyzed, involving 32 males and 13 females, aged 2-14 years.Among them, 27 cases were simple bone cyst, 15 cases were fibrous dysplasia, 3 cases were Langerhans histiocytosis, and 17 cases were accompanied with pathological fracture.Patients without pathological fracture were divided into 2 groups according to different surgical methods: lesion removal and bone grafting+ LCH-PHP preventive fixation group (group A) and lesion removal and bone grafting group (group B). Patients with pathological fracture were divided into 2 groups according to the different surgical methods: lesion removal and bone grafting+ LCH-PHP fixation and reduction group (group C) and lesion removal and bone grafting group (group D). The loss of femoral neck-stem angle, healing time and Musculoskeletal Tumor Society (MSTS) scores were compared between group A and group B. Changes of femoral neck-stem angle were compared between group C and group D at postoperative 1 day and the last follow-up.Results:Forty-five patients were followed up for an average of 2.6 (1-4) years.The loss of femoral neck-stem angle in group A and group B were (0.50 ± 0.45)°, and (2.26 ± 1.64)°, which was significantly different ( t=-3.193, P<0.05). There were no significant differences in the healing condition, healing time and MSTS scores at the last follow-up between group A and group B(all P>0.05). In group C, the ipsilateral femoral neck-stem angle at the postoperative 1 day and the last follow-up were (133.67±4.53)°and (133.00±4.32)°, respectively, which was comparable ( P>0.05). In group D, the ipsilateral femoral neck-stem angle at the postoperative 1 day and the last follow-up were (133.36±7.82)°and (130.63±8.24)°, respectively, which was significantly different ( t=7.11, P<0.05). Conclusions:LCH-PHP fixation can effectively maintain the femoral neck-stem angle and prevent the occurrence of pathological fracture children with Erol Ⅱ-Ⅳ tumor-like lesions of femoral neck.

Essential tremor (ET) is one of the most common movement disorders. Its clinical manifestations not only include typical kinetic and/or postural tremors, but also other non-motor symptoms such as cognitive dysfunction, sleep disturbance, and dysosmia. The exact etiology and pathogenesis of ET is still unknown. Approximately 60% of ET patients have a family history, and genetic factor plays an important role in the onset of the disease. Researchers have so far identified 3 genetic loci (ETM 1-3) through family studies, and proposed additional causative genes such as FUS, HTRA2, TENM4, NOS3 and susceptibility genes such as LINGO, SLC1A2, and GABA. This review focuses on the progress made in genetic research on ET.
Essential Tremor ; etiology ; genetics ; Genetic Predisposition to Disease ; Genetic Research ; High-Temperature Requirement A Serine Peptidase 2 ; genetics ; Humans ; Membrane Proteins ; genetics ; Nerve Tissue Proteins ; genetics ; RNA-Binding Protein FUS ; genetics

Objective To investigate clinical effects differences of ropivacaine combined with two doses of sufentanil on labor analgesia.Methods300 lying-in women with painless childbirth in the period from January 2016 to December 2016 in department of obstetrics in Deqing hospital of traditional Chinese medicine were chosen and randomly divided into two groups including A group (150 cases) with ropivacaine combined with conventional doses of sufentanil for 7.5μg and B group (150 cases) with ropivacaine combined with low doses of sufentanil for 2.5μg;the block onset time of sensory and motor, arrival time at the maximum block level, modified Bromage score, Apgar score, VAS score at different time points and adverse reaction rate of both groups were compared.ResultsTwo groups of maternal sensory and motor block work time and reach the highest block plane comparative differences had no statistical significance.The modified Bromage score of B group were significant better than A group(P<0.05).The adverse reaction rate of B group were significant lower than A group(P<0.05).ConclusionCompared with conventional dosage sufentanil, luo pp paid in combination with low dose used in epidural sufentanil analgesia can obviously reduce the working time, improve the quality of anesthesia and reduce the incidence of adverse reactions, is suitable for clinicians to choose application.

Objective To analyze the clinical characteristics and treatment strategy of ankle supination fracture in children.Methods From January 2012 through July 2014,89 children were treated at our department for ankle fracture caused by supination sprain according to their medical history,physical examination,X-ray films and CT three-dimensional reconstruction of the ankle.Appropriate protocols were applied on the basis of Lauge-Hansen classification,type and displacement of their fractures.Of them,52 belonged to the supination-adduction type (the extramalleolus fracture was of Odgen type Ⅶ in 51 children whose epiphyseal plate of distal fibula had not been closed),35 to the supination-extorsion type (32 cases had tri-plane fracture and 3 Tillaux fracture),and the remaining 2 did not fit the Lauge-Hansen classification.Surgical treatment was applied in 32 cases and conservative treatment in 57 ones.Results All the children received outpatient follow-up from 12 to 24 months(mean,18 months).No bone nonunion,osteoarthritis,or fracture malunion was found.The American Orthopaedic Foot & Ankle Society scores averaged 92 points (range,from 88 to 100 points) at the last follow-ups.Conclusions In children whose epiphyseal plate is nearly closed,supination-adduction sprain likely causes an extramalleolus fracture of Salter-Harris type Ⅰ or type Ⅱ,but in children whose epiphyseal plate is unclosed,an epiphyseal fracture of Odgen type Ⅶ is inclined to happen.A Tillaux fracture or tri-plane fracture at the level of distal tibiofibular syndesmosis results often from supination-extorsion sprain in children.For fractures involving epiphysis or epiphyseal plate,anatomical reduction and proper fixation are critical to functional recovery and reducing complications.