Association of sugar-sweetened beverage consumption and sleep quality with anxiety symptoms: a cross-sectional study of Tibetan college students at high altitude.

Background: Research on the association between sugar-sweetened beverages (SSBs) consumption and sleep quality with anxiety symptoms has been highly emphasized. However, past studies have focused on college students in plains areas, while fewer research studies have been conducted on Tibetan college students at high altitudes. Whether this association changes due to ethnicity or altitude is unclear. The present study will contribute to the prevention and intervention of depressive symptoms among Tibetan college students at high altitude. Methods: A self-assessment questionnaire was administered to 3,026 university students (1,491 boys students, 49.27%) on SSBs consumption, sleep quality and anxiety symptoms status in the Tibetan Plateau, a high-altitude region of China. Logistic regression analysis and ordered logistic regression analysis in generalized linear model were used to analyze the association between SSBs consumption and sleep quality with anxiety symptoms. Results: The prevalence of anxiety symptoms among Tibetan college students at high altitude was 26.9%. SSBs consumption of ≤1 times/week, 2-5 times/week, and ≥ 6 times/week were 20.7, 28.1, and 45.7%, respectively, with statistically significant differences (χ2 value of 134.353, p < 0.001). Anxiety detection rates for Sleep quality of Good (PSQI ≤5), Moderate (PSQI 6-7), and Poor (PSQI >7) were 16.8, 19.8, and 32.0%, respectively, and the difference was also statistically significant (χ2 value was 73.761, p < 0.001). The ordered logistic regression analysis in the generalized linear model showed that, overall, the group of college students with SSBs ≤1 times/week and sleep quality of Good served as the reference group, and the group with SSBs ≥6 times/week and sleep quality of Poor (OR: 5.06, 95% CI: 3.75-6.83) had the highest risk of anxiety symptoms. Conclusion: SSBs consumption and sleep quality were associated with anxiety symptoms, and there was an interaction effect. Effective control of SSBs consumption and improvement of sleep quality may be important factors in preventing and reducing the occurrence of anxiety symptoms.

UBR4 deficiency causes male sterility and testis abnormal in Drosophila.

Introduction: It has been established that UBR4 encodes E3 ubiquitin ligase, which determines the specificity of substrate binding during protein ubiquitination and has been associated with various functions of the nervous system but not the reproductive system. Herein, we explored the role of UBR4 on fertility with a Drosophila model. Methods: Different Ubr4 knockdown flies were established using the UAS/GAL4 activating sequence system. Fertility, hatchability, and testis morphology were studied, and bioinformatics analyses were conducted. Our results indicated that UBR4 deficiency could induce male sterility and influent egg hatchability in Drosophila. Results: We found that Ubr4 deficiency affected the testis during morphological analysis. Proteomics analysis indicated 188 upregulated proteins and 175 downregulated proteins in the testis of Ubr4 knockdown flies. Gene Ontology analysis revealed significant upregulation of CG11598 and Sfp65A, and downregulation of Pelota in Ubr4 knockdown flies. These proteins were involved in the biometabolic or reproductive process in Drosophila. These regulated proteins are important in testis generation and sperm storage promotion. Bioinformatics analysis verified that UBR4 was low expressed in cryptorchidism patients, which further supported the important role of UBR4 in male fertility. Discussion: Overall, our findings suggest that UBR4 deficiency could promote male infertility and may be involved in the protein modification of UBR4 by upregulating Sfp65A and CG11598, whereas downregulating Pelota protein expression.

Efficient strategies based on behavioral and electrophysiological methods for epilepsy-related gene screening in the Drosophila model.

Introduction: With the advent of trio-based whole-exome sequencing, the identification of epilepsy candidate genes has become easier, resulting in a large number of potential genes that need to be validated in a whole-organism context. However, conducting animal experiments systematically and efficiently remains a challenge due to their laborious and time-consuming nature. This study aims to develop optimized strategies for validating epilepsy candidate genes using the Drosophila model. Methods: This study incorporate behavior, morphology, and electrophysiology for genetic manipulation and phenotypic examination. We utilized the Gal4/UAS system in combination with RNAi techniques to generate loss-of-function models. We performed a range of behavioral tests, including two previously unreported seizure phenotypes, to evaluate the seizure behavior of mutant and wild-type flies. We used Gal4/UAS-mGFP flies to observe the morphological alterations in the brain under a confocal microscope. We also implemented patch-clamp recordings, including a novel electrophysiological method for studying synapse function and improved methods for recording action potential currents and spontaneous EPSCs on targeted neurons. Results: We applied different techniques or methods mentioned above to investigate four epilepsy-associated genes, namely Tango14, Klp3A, Cac, and Sbf, based on their genotype-phenotype correlation. Our findings showcase the feasibility and efficiency of our screening system for confirming epilepsy candidate genes in the Drosophila model. Discussion: This efficient screening system holds the potential to significantly accelerate and optimize the process of identifying epilepsy candidate genes, particularly in conjunction with trio-based whole-exome sequencing.

Evaluation of RT-LAMP Assay for Rapid Detection of SARS-CoV-2.

OBJECTIVE: To evaluate the accuracy of the reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in community or primary-care settings. METHOD: We systematically searched the Web of Science, Embase, PubMed, and Cochrane Library databases. We conducted quality evaluation using ReviewManager software (version 5.0). We then used MetaDisc software (version 1.4) and Stata software (version 12.0) to build forest plots, along with a Deeks funnel plot and a bivariate boxplot for analysis. RESULT: Overall, the sensitivity, specificity, and diagnostic odds ratio were 0.79, 0.97, and 328.18, respectively. The sensitivity for the subgroup with RNA extraction appeared to be higher, at 0.88 (0.86-0.90), compared to the subgroup without RNA extraction, at 0.50 (0.45-0.55), with no significant difference in specificity. CONCLUSION: RT-LAMP assay exhibited high specificity regarding current SARS-CoV-2 infection. However, its overall sensitivity was relatively moderate. Extracting RNA was found to be beneficial in improving sensitivity.

H3K36me2 methyltransferase NSD2 orchestrates epigenetic reprogramming during spermatogenesis.

Spermatogenesis is precisely controlled by sophisticated gene expression programs and is driven by epigenetic reprogramming, including histone modification alterations and histone-to-protamine transition. Nuclear receptor binding SET domain protein 2 (Nsd2) is the predominant histone methyltransferase catalyzing H3K36me2 and its role in male germ cell development remains elusive. Here, we report that NSD2 protein is abundant in spermatogenic cells. Conditional loss of Nsd2 in postnatal germ cells impaired fertility owing to apoptosis of spermatocytes and aberrant spermiogenesis. Nsd2 deficiency results in dysregulation of thousands of genes and remarkable reduction of both H3K36me2 and H3K36me3 in spermatogenic cells, with H3K36me2 occupancy correlating positively with expression of germline genes. Nsd2 deficiency leads to H4K16ac elevation in spermatogenic cells, probably through interaction between NSD2 and PSMA8, which regulates acetylated histone degradation. We further reveal that Nsd2 deficiency impairs EP300-induced H4K5/8ac, recognized by BRDT to mediate the eviction of histones. Accordingly, histones are largely retained in Nsd2-deficient spermatozoa. In addition, Nsd2 deficiency enhances expression of protamine genes, leading to increased protamine proteins in Nsd2-deficient spermatozoa. Our findings thus reveal a previously unappreciated role of the Nsd2-dependent chromatin remodeling during spermatogenesis and provide clues to the molecular mechanisms in epigenetic abnormalities impacting male reproductive health.

Efficacy of three-dimensional guide plate technique guided sacral 2 alar iliac screws fixation in patients with degenerative kyphoscoliosis.

OBJECTIVE: To investigate the efficacy of three-dimensional (3D) guide plate technique guided sacral 2 alar iliac (S2AI) screws fixation in patients with degenerative kyphoscoliosis. METHODS: Eighty-four patients with degenerative kyphoscoliosis who were admitted to our hospital were selected as the subjects. They were divided into control group and observation group by the random number table method, with 42 patients in each group. S2AI free-hand screw implantation technique was adopted for screws fixation in control group; 3D guide plate technique guided S2AI screw implantation was used for screws fixation in observation group. A 2-year routine follow-up was carried out after the surgery. The followings were compared: screw parameters of preoperative pre-set screw trajectory and postoperative actual screw trajectory: sagittal angle (SA), transverse angle (TA), horizon distance from the entry point to the median sacral crest (HD), vertical distance from the entry point to the superior margin of the second posterior sacral foramina (VD), and the incidence rate of complications; scoliosis Cobb angle, sagittal vertical axis, C7 plumb line-center sacral vertical line (C7PL-CSVL), regional kyphosis Cobb angle (RK), pelvic incidence (PI), and pelvic tilt (PT) before, after and 2 years after surgery; Oswestry disability index (ODI) and shot form 36 health survey questionnaire (SF-36) before and 2 years after surgery. RESULTS: The difference between preoperative simulation and postoperative actual values of SA, TA, HD and VD was significantly lower in observation group than in control group (P<0.001). Scoliosis Cobb angle, lumbar lordosis, C7PL-CSVL, RK, PI and PT after and 2 years after surgery were significantly improved than those before surgery in the two groups (P<0.001), and there was no significant difference between those after surgery and 2 years after surgery (P>0.05). Patients in the two groups had significantly lower ODI scores and higher SF-36 scores at 2 years after surgery than those before surgery (both P<0.001), and there was no significant difference at 2 years after surgery between the two groups (P>0.05). The incidence rate of complications in observation group was significantly smaller than that in control group (P<0.05). CONCLUSION: 3D guide plate technique guided S2AI fixation can significantly increase the accuracy of screw implantation, effectively correct degenerative kyphoscoliosis, achieve rigid internal fixation, improve patient's spinal function and quality of life, and greatly enhance surgical safety, which is worthy of clinical popularization.

Dynamic pattern of histone H3 core acetylation in human early embryos.

After fertilization, highly differentiated sperm and oocyte are reprogrammed to totipotent embryo, which subsequently cleavages and develops into an individual through spatial-temporal differentiation. Histone modifications play critical roles to coordinate with other reprogramming events in early stages of embryogenesis. However, most of studies focus on modifications at N-terminus of histones, those at nucleosome core were not well understood. Here, we characterize the three key acetylation events in the histone H3 core, H3K56/K64/K122ac, in early human embryos. The three residues localize at DNA entry-exit position of the nucleosome. Globally, H3K56ac, H3K64ac and H3K122ac were detectable throughout preimplantation stages, with H3K64ac levels being relatively stronger and H3K122ac levels being much weaker. Besides, H3K56ac level had a peak at two-cell stage. Moreover, we found that LINEs also peak at two-cell stage, and H3K56ac was enriched at young LINE-1 in human ESCs, supporting that H3K56ac is an important driving force for young LINE-1 activation in human preimplantation embryos. Our results suggest that acetylation in the nucleosome core of histone H3 is dynamic and various during preimplantation development, and may drive diverse chromatin remodeling events in this developmental window.

Rybp orchestrates spermatogenesis via regulating meiosis and sperm motility in mice.

Ring1 and Yin Yang 1-Binding Protein (RYBP) is a member of non-canonical polycomb repressive complex 1 to mediate monoubiquitination of histone H2A at lysine 119. It plays an important role in development, but its role in reproduction remains illusive. In this study, we used Rybp conditional knockout mouse model to genetically ablate Rybp in male germ cells. We found that Rybp deficiency during spermatogenesis led to smaller testes, loss of germline cells, disturbed meiosis, increased apoptosis of spermatocytes, decreased sperm motility, and reduced global H3K9me3, without impacting retrotransposon expression. Meanwhile, we depleted Rybp during oogenesis, but oocyte maturation and preimplantation development were normal. Our findings demonstrate that RYBP plays important roles in spermatogenesis through regulating meiosis and sperm motility.

Generation of an iPSC line (HUSTi002-A) from fibroblasts of a patient with Sertoli cell-only syndrome carrying c.731_732delAT in PIWIL2 gene.

Sertoli cell-only syndrome (SCOS) is a severe phenotype of male infertility; autosomal gene defects are thought to be the causes for this disease. The iPSC line generated from a SCOS patient carrying a mutation in PIWIL2 gene expresses pluripotent markers, has a normal karyotype and the mutation c.731_732delAT in PIWIL2 gene and is able to differentiate into three germ layers. This cell line will help to study the pathogenesis of SCOS, and the roles of PIWIL2 in human germ cells development and spermatogenesis.

H3.3 impedes zygotic transcriptional program activated by Dux.

During development, fertilization triggers totipotency establishment, featured by zygotic genome activation/embryonic genome activation (ZGA/EGA). Mouse embryonic stem cells (mESCs) occasionally cycle through a two-cell (2C)-like status with activated expression of Dux and its targeted ZGA genes. Here, we demonstrate that deficiency of histone variant H3.3 dramatically stimulates expression of ZGA genes in mESCs. Our analysis revealed that H3.3 directly associates with Dux locus and inhibits Dux expression, therefore it is an important upstream regulator of Dux. Our finding is further supported by transcriptome change in early mouse embryos with H3.3 knockdown. We suggest that proper H3.3 level in early embryos is important to orchestrate ZGA activity for totipotency establishment.

Accuracy and Safety of Lateral Vertebral Notch-Referred Technique Used in Subaxial Cervical Pedicle Screw Placement.

BACKGROUND: Biomechanical studies revealed that pedicle screw instrumentation has a superior stabilizing effect compared with other internal fixations in reconstructing the subaxial cervical spine. However, severe neurovascular risks preclude surgeons from routinely conducting pedicle screw manipulation in cervical spine. OBJECTIVE: To evaluate the accuracy and safety of the lateral vertebral notch (LVN)-referred technique used in subaxial cervical pedicle screw (CPS) placement. METHODS: One hundred thirty-five consecutive retrospective patients with cervical disorders underwent the LVN-referred technique for CPS placements in 3 spine centers. Postoperative pedicle perforations were confirmed by CT scans to assess the technical accuracy. Neurovascular complications derived from CPS misplacements were recorded to evaluate the technical safety. RESULTS: A total of 718 CPSs were inserted into subaxial cervical spine. Postoperative CT scans revealed that the accuracy of CPS placement was superior. Neither vertebral artery injury nor spinal cord injury occurred. One radiculopathy was from a unilateral C6 nerve root compression. A screw-related neurovascular injury rate of 0.7% occurred in this cohort. Additionally, there was no significant difference in the accuracy of CPS placement among 3 surgeons (H = 1.460, P = .482). The relative standard deviation values revealed that technical reproducibility was acceptable. Furthermore, there was no significant difference between the patients' pedicle transverse angles and inserted CPS transverse angles from C3 to C7 (all P > .05). CONCLUSION: The LVN is a reliable and consistent anatomic landmark for CPS placement. The accuracy and safety of subaxial CPS placement by using LVN-referred technique are highly acceptable, which may endow this technique to be practicably performed in selected patients.

Expert consensus for PVCR in severe, rigid and angular spinal deformity treatment: The Kunming consensus.

In the past decades, an increasing number of surgeons started using posterior vertebral column resection (PVCR) to treat severe, rigid and angular spinal deformities. Little high-level evidence is available to guide surgical treatment. The aims of our study were to identify important surgical strategies and key technical points of Chinese experts who have extensive experience in the management of severe, rigid and angular spinal deformities using PVCR, and to standardize and unify the current core concepts. Workgroups of consensus were formed by selecting nationwide representing experts and comprehensive consultations. Eight task forces for major issues were established, then retrieval of literature, collection of expert opinions and writing of review articles were carried out. A modified Delphi process was chosen in round-table forum with three face-to-face meetings. Consensus was reached with items graded more than seven points including: indications and contraindications of PVCR; review PVCR in the evolution of spinal osteotomies; The corrective mechanism and safety of spinal cord; monitoring and responses of spinal cord crisis; characteristics and therapeutic outcome of pulmonary function; management of bleeding during PVCR; relationship of pedicle screw insertion and spinal cord safety; and analysis of non-neurologic complications and prevention strategies. In conclusion, The essential properties regarding PVCR procedure are tightly linked with various factors such as medical and surgical indication, range and level of vertebral column resection, strategies of correction, corrective efficiency and control of neurological risk. PVCR is used mainly for severe, rigid spinal deformity that is not manageable by other osteotomy techniques.

[Ponte OSTEOTOMY FOR OLD OSTEOPOROTIC MULTI-SEGMENT VERTEBRAL FRACTURE COMBINED WITH THORACOLUMBAR KYPHOSIS IN ELDERLY].

OBJECTIVE: To investigate the clinical outcome of Ponte osteotomy for old osteoporotic multi-segment vertebral fracture combined with thoracolumbar kyphosis in elderly. METHODS: Between April 2007 and December 2012, 11 elderly patients with thoracolumbar kyphosis caused by old osteoporotic multi-segment vertebral fracture received Ponte osteotomy. There were 4 males and 7 females, with an average age of 69 years (range, 62-76 years) and with an average disease duration of 4 years (range, 10 months to 7 years). A total of 51 vertebrae was involved in fracture, including T5 in 1, T8 in 1, T9 in 1, T10 in 5, T11 in 8, T12 in 11, L1 in 11, L2 in 10, and L3 in 3. Based on Frankel classification, there were 9 cases of grade E and 2 cases of grade D. The Cobb angle of thoracolumbar kyphosis and the distance of sagital plane between C7 plump line and the posterior superior corner of S1 (sagittal vertical axis distance, SVA) were measured at pre-operation, at 2 weeks after operation, and last follow-up. The effectiveness of the treatment was appraised by visual analogue scale (VAS). RESULTS: All the operations were successfully completed. Cerebrospinal fluid leakage occurred in 3 cases, and was cured after symptomatic treatment. Eleven patients were followed up 22 months on average (range, 7-38 months). Back pain symptoms, the appearance, and the trunk balance were significantly improved after operation. Postoperatively neurological symptoms were not aggravated. In 2 cases of grade D, 1 case recovered to grade E, and 1 case had no improvement. Osseous fusion was observed at 6.7 months on average (range, 5-9 months) in 10 patients except 1 patient who had no complete fusion at 7 months after operation. There was no infection, failure of internal fixation, or other complications. At 2 weeks after operation and last follow-up, the VAS score, Cobb angle, and SVA were significantly improved when compared with preoperative ones(P < 0.05), but no significant difference was found between at 2 weeks and last follow-up (P > 0.05). The correction rate of Cobb angle was 70.6% ± 2.2% at 2 weeks after operation and was 66.7% ± 1.3% at last follow-up, showing no significant difference (t = 25.20, P = 0.13). CONCLUSION: Ponte osteotomy is effective for old osteoporotic multi-segment vertebral fracture combined with thoracolumbar kyphosis in elderly. It can obtain satisfactory sagittal balance.