Humans ; Gastrointestinal Microbiome ; Constipation/therapy* ; Probiotics/therapeutic use* ; Patients

OBJECTIVE To develop a high-throughput screening assay for the discovery of Omicron variant main protease(OM-Mpro) inhibitors based on the principle of fluorescence resonance energy transfer(FRET). METHODS The recombinant OM-Mpro enzyme was expressed in Escherichia coli Rosetta(DE3) cells, and further purified by a HisTrapTM chelating column. Subsequently, the enzymatic activity of OM-Mpro and wild type main protease(WT-Mpro) enzymes and inhibition of nirmatrelvir against both proteases were measured using FERT assay. With the FRET assay, OM-Mpro inhibitors were identified via high-throughput screening of an approved drug library. RESULTS The active OM-Mpro enzyme was successfully prepared from E. coli cells. OM-Mpro and WT-Mpro enzymes possessed the same enzymatic activity, and OM-Mpro remained susceptible to nirmatrelvir in vitro. Through high-throughput screening of the marketed drug library, it was found that cetylpyridinium chloride(CPC) is a mixed-type OM-Mpro inhibitor in vitro with an IC50 value of 8.76 μmol·L−1. CONCLUSION A robust FRET assay has been successfully developed based on the production of active OM-Mpro enzyme for screening of its inhibitors, and CPC is identified as a potential lead compound against OM-Mpro in vitro. This study provides a promising avenue for rapid discovery of broad-spectrum antivirals against coronavirus protease.

Objective To observe the changes in the subfoveal choroidal thickness(SFCT)of children and adoles-cents with different refractive states using optical coherence tomography angiography.Methods A total of 171 children and adolescents were followed.They were divided into the lower primary school group(6-8 years old),upper primary school group(9-11 years old),and junior high school group(12-14 years old)according to their age at the time of en-rollment.Dioptric examinations(including best corrected visual acuity,diopter,intraocular pressure,corneal curvature,axial length and SFCT)were performed,data collection was conducted twice in half a year(initial examination and review after half a year),and the eyeball parameters and changes in eyeball parameters after half a year among all groups were compared.Results The axial length and SFCT of subjects had significant differences among all groups(both P＜0.05).In children and adolescents,the axial length gradually lengthened and SFCT gradually thickened with age,while intraocular pressure and corneal curvature were not associated with age(both P＞0.05).In the initial examination and review after half a year,there was no significant difference in intraocular pressure,corneal curvature and SFCT of subjects with differ-ent refractive states in all groups(all P＞0.05),while the axial length of myopic subjects was greater than that of non-my-opic subjects in all groups(all P＜0.05).In the review after half a year,the SFCT of non-myopic subjects in the lower pri-mary school group and upper primary school group was significantly thickened(P＜0.001,P=0.003),while there was no significant difference in SFCT of myopic subjects in all groups compared with the value half a year ago(all P＞0.05).The axial length of all subjects showed a positive correlation with the SFCT in the initial examination and review after half a year(r=0.354,0.228,P＜0.05).Conclusion Myopia affects the increase in SFCT in children and adolescents.

This study aims to investigate the function of lmo2363 gene in stress resistance of Liste-ria monocytogenes strain LM83-1.In this study,the lmo2363 gene deletion strain and complement-ation strain of Listeria monocytogenes were constructed using overlapping extended PCR and ho-mologous recombination techniques,and the growth ability,stress survival rate and biofilm forma-tion ability of wild,deletion strain and complementation strain were compared under different stress environments.lmo2363 gene deletion strain and complementation strain of Listeria monocy-togenes were successfully constructed in this experiment.The growth curves showed that the growth capacity of the deletion strain was weaker than the wild strain LM83-1 under 4 ℃,7％NaCl,10％NaCl,3.5％ethanol,4.0％ethanol and pH5 stress(P＜0.001).The results of stress survival test showed that the survival rate of the deletion strain was significantly lower than the wild strain after 1 h treatment with pH3 and 10 mmol/L H2 O2 stress(P＜0.010).The biofilm forming ability of the deletion strain was decreased compared with that of the wild strain(P＜0.050).This study confirmed that lmo2363 gene mediated the adaptation of LM to low temperature,high osmotic pressure,ethanol and acid stress environment and affected the formation of LM bio-film.This study laid a foundation for further exploring the function of lmo2363 gene in the stress resistance process of Listeria monocytogenes.

Objective:To propose a method to measure the femoral anteversion angle based on the three-dimensional model of femur.Methods:From January 2019 to December 2023, a total of 50 patients (100 femurs) with patellar instability who underwent full-length CT examination of both lower limbs in Tianjin Hospital were retrospectively analyzed, including 16 males and 34 females, aged 28.5±11.9 years (range, 16-57 years). All patients underwent CT examination of both lower limbs, and the imaging data were imported into Mimics 21.0 software as DICOM format files. The femur was extracted by the threshold segmentation function, and the 3D model of the femur was reconstructed by appropriate trimming and smoothing. Three-dimensional models of the femur from 30 patients (60 femurs) were selected and imported into 3-matic 13.0 software for labeling of anatomical points and drawing of axes. The angle between the axis of the distal femur and the axis of the proximal femur was the femoral anteversion angle. Two observers positioned the spatial coordinates of the intercondylar fossa apex, the greater trochanter apex, the center of the femoral head, the femoral through-condylar axis and the posterior condylar tangent marker points on a 3D model of the femur to compare the differences between the groups. The femoral anteversion angles measured by different three-dimensional measurement methods were compared with the clinically reported femoral anteversion angles.Results:The difference between the femoral head center and intercondylar fossa apex coordinate data of 30 patients (60 femurs) measured by the two observers was not statistically significant ( P>0.05), whereas the data for the Y-axis coordinate of the greater trochanter apex were 25.77±23.21 and 22.08±25.73, respectively, and the difference was statistically significant ( t=2.906, P=0.017). The difference between the data of femoral through condylar and posterior condylar tangential coordinates of 30 patients (60 femurs) measured by the two observers was not statistically significant ( P>0.05). Using the femoral mechanical axis as the projection direction, the angle between the transcondylar axis and the posterior condylar tangent line was 5.97°±2.02° (range, 1.48°-12.08°). The anterior tilt of the femur measured by the two observers in the femoral neck cylinder fitting method was 23.35°±7.45° and 24.94°±9.01°, respectively, and the difference was statistically significant ( t=-2.147, P=0.040). The anterior inclination of the femur measured by the two observers in the femoral head ball enlargement method was 24.63°±7.66° and 25.12°±8.84°, the difference was not statistically significant ( t=-0.820, P=0.419). Using the femoral head ball expansion method to locate the proximal femoral axis, two observers measured the anterior tilt angle of the femur in 50 patients (100 femurs) according to the method described above, which was 23.30°±9.41° and 23.57°±9.97°, respectively, without a statistically significant difference ( t=-0.664, P=0.508), both smaller than the 29.56°±8.74° measured based on two-dimensional images in the imaging report, and the difference was statistically significant ( P<0.05). Conclusion:The femoral anteversion angle measured by the angle between the axis of the femoral condyle and the axis of the femoral neck based on the three-dimensional model is accurate and effective, and the measured value is smaller than that based on the two-dimensional images in clinical practice.

Objective:To observe the effect of shift work on the stability of the circadian clock and insulin sensitivity in non-overweight/obese individuals with normal blood glucose, and explore underlying connection.Methods:Female shift working nurses in the Department of Blood Transplantation and non-shift working nurses in the Health Management Center in the First Affiliated Hospital of Zhengzhou University were divided into shift worker group (SW group) and non-shift worker group (NSW group). Serum inflammatory factors [interleukin-6 (IL-6), tumor necrosis factor-α(TNF-α)], adipokines (adiponectin, leptin, chemerin, visfatin), and melatonin levels were measured using enzyme linked immunosorbent assay (ELISA). Realtime fluorescence quantitative PCR was performed to detect peripheral blood circadian clock genes circadian locomotor output cycles protein kaput(Clock) and brain and muscle ARNT-like protein 1(Bmal1). Cortisol and fasting insulin were measured by chemiluminescent microparticle immunoassay, and HbA 1C was measured by capillary electrophoresis. In addition, visceral fat area (VFA) was assessed with bioelectrical impedance analyzer, and mid-sleep time composite phase deviations (CPD) was calculated based on the International Physical Activity Short Questionnaire. Results:SW group had lower serum level of melatonin ( P=0.023) and higher cortisol ( P=0.001) than the NSW group, and altered mRNA expression of Clock and Bmal1 ( P=0.034, P=0.047). Fasting blood glucose and HbA 1C in the SW group, although in the normal range, had been higher than in the NSW group ( P=0.011, P=0.033). Although body mass index was normal in SW group, VFA had been higher than that of the NSW group ( P=0.010). And homeostasis model assessment for insulin resistance (HOMA-IR), IL-6, TNF-α, leptin, chemerin, and visfatin were significantly higher in the SW group than NSW group ( P=0.033, P=0.012, P=0.001, P=0.011, P=0.021, P=0.007). In addition, adjusting for body mass index and activity factors revealed a significant positive correlation between CPD and VFA ( r=0.434, P=0.049), inflammatory factors IL-6 ( r=0.514, P=0.017) and TNF-α ( r=0.700, P<0.001) and pro-inflammatory adipokines leptin ( r=0.473, P=0.030), chemerin ( r=0.439, P=0.047), visfatin ( r=0.521, P=0.015). Conclusion:Shift work can affect circadian clock, with increased visceral adiposity, pro-inflammatory adipokines, inflammatory factors and decreased insulin sensitivity in women without overweight/obese.

Objective:To analyze nine cases of pregnancy and lactation-associated osteoporosis(PLO) along with a literature review to improve clinicians′ knowledge of the disease.Methods:We collected medical history, laboratory examination, bone mineral density(BMD) scan, treatment and follow-up data of 9 inpatients with PLO in the Endocrinology Department of the First Affiliated Hospital of Zhengzhou University from January 2014 to June 2021.Results:The median age of onset of 9 patients with PLO was 30 years(22-37 years). All 9 patients presented low back pain, and the median time of low back pain after childbirth was 2.5 months(0.5-7 months). 89% of the patients presented vertebral compression fractures, and the mean number of vertebral fractures was 4.6. Eight patients with PLO showed osteoporosis by dual X-ray absorpiometry(DXA) scan, and one patient showed bone loss by quantitative CT scan. Osteoporosis predominated in the trabecular bone. After the diagnosis, nine patients with PLO stopped breastfeeding and were given calcium and vitamin D preparations. Seven patients were given bisphosphonates and one patient was treated with teriparatide followed by denosumab. The back pain of all patients was relieved, with the median of relief time being 3 months(7 d-6 m). After treatment initiation, BMD was increased in eight patients, especially obvious in the lumbar spine. No new clinical fractures occurred during the follow-up.Conclusions:For women with low back pain and shortened height in late pregnancy or breastfeeding, the possibility of PLO should be considered. Prompt diagnosis and early medical interventions are of utmost importance to reduce the risk of subsequent fractures and improve the prognosis.

Objective:To explore the accuracy and repeatability of the smartphone scoliosis screening APP developed by our team in the measurement of the Cobb angle of adolescent idiopathic scoliosis (AIS).Methods:Clinical data of 60 patients with AIS admitted to Department of Spine and Spinal Surgery, Henan Province People′s Hospital from August 2020 to February 2021 were analyzed retrospectively.Three surveyors measured the coronal main curvature Cobb angle, sagittal thoracic kyphosis (TK) angle, thoracolumbar kyphosis (TLK) angle and lumbar kyphosis (LL) angle on whole-spine lateral X-ray films of 60 patients with AIS by means of the protractor, scoliosis screening APP and computerized Picture Archiving and Communication System (PACS). The time and results of each measurement were recorded.The measurement was repeated once after 2 weeks.Paired t-test was used to compare the measurement time of the APP method and the protractor method.Taking the Cobb angle measurement results of the PACS system as the reference standard, the accuracy of Cobb angle measurement by the APP method was analyzed by paired t-test.The repeatability of the surveyor and the consistency between the surveyors was compared by intraclass correlation coefficient (ICC). Results:Among the 60 patients with AIS, there were 17 males and 43 females, aged from 10 to 16 years [(12.2±2.4) years]. The main bends were thoracic curvature (Lenke Ⅰ) in 23 cases, Lenke Ⅱ in 18 cases and thoracolumbar curvature/lumbar curvature (Lenke V) in 19 cases.The APP method took significantly less time to measure the Cobb angle than the protractor method ( P<0.05). There was no significant difference in the Cobb angle measured by the APP method and PACS method ( P>0.05). The results of the coronal main curvature Cobb angle, TK angle, TLK angle and LL angle measured by 3 surveyors through the APP method were all in good agreement (ICC=0.990, 0.988, 0.986, 0.987). The repeatability (ICC 0.973-0.982) of the coronal main curvature Cobb angle, TK, TLK and LL measured twice before and after the APP method were both better that of the protractor method (ICC 0.933-0.954). Conclusions:Compared with the traditional protractor, the smartphone scoliosis screening APP has the advantages of short measurement time, high efficiency, excellent accuracy and good repeatability in measuring the Cobb angle of AIS.

Objective:To analyze the postoperative paravertebral muscle degeneration and its correlations with health related quality of life (HRQL) in patients undergoing minimally invasive surgery-transforaminal lumbar interbody fusion (MIS-TLIF).Methods:The clinical data of the 50 patients were retrospectively analyzed who had undergone single-segmental MIS-TLIF at Department of Spinal Cord Surgery, The People's Hospital of Henan Province from January 2019 to December 2021. The relative volumes of lumbar posterior muscle (LM), the relative volumes of the psoas major (PM), and the rates of fatty degeneration (FD) of the fused segment and its adjacent segments were compared respectively between preoperation, 6 and 12 months postoperation. The correlations were analyzed between the HRQL scores [visual analog scale (VAS) for pain and Oswestry disability index (ODI)] and the relative LM volumes, the relative PM volumes, and the FD rates of the fused segment and its adjacent segments at 12 months postoperation.Results:Compared with the preoperative values, the relative LM volumes and the relative PM volumes of the fused segment and its adjacent segments at 6 and 12 months postoperation were significantly reduced while the FD rates significantly increased. However, the FD rate of the fused segment at 12 months postoperation (20.6% ± 6.1%) was significantly lower than that at 6 months postoperation (29.7% ± 8.2%) ( P < 0.05). The VAS score was strongly negatively or positively correlated with the relative LM volume ( r = -0.819, P < 0.001) and the FD rate ( r = 0.86, P < 0.001) of the fused segment, and moderately negatively correlated with the relative PM volume ( r = -0.435, P = 0.016). The ODI index was moderately negatively correlated with the relative LM volume ( r = -0.512, P = 0.004) and the relative PM volume ( r = -0.402, P = 0.020) of the fused segment, but moderately positively correlated with the FD rate of the fused segment ( r = 0.565, P = 0.001). There was a moderate negative correlation between the ODI index and the relative LM volume of the adjacent segments ( r = -0.478, P = 0.012). Conclusions:After MIS-TLIF, the volume of the paravertebral muscles decreases and the dorsal muscles develop fatty degeneration. The improvement of LM fatty degeneration may be observed by 12-month follow-up in the fused segment, but not in the adjacent segments. The LM volume and the FD rate of the fused segment are the most closely related to the postoperative HRQL.

OBJECTIVE: To report on the diagnosis and treatment process and clinical characteristics of a child with disorder of sex development (DSD) and to conduct pathological, imaging and genetic analysis for the patient. METHODS: Clinical data of the patient were collected. Genetic testing including chromosomal karyotyping, fluorescence in situ hybridization (FISH), copy number variations (CNVs) analysis, SRY gene detection and multiple ligation-dependent probe amplification (MLPA) were carried out. RESULTS: The patient had a social gender of male, with a history of hypospadia and breast development. Sex hormone tests showed slightly raised prolactin. Imaging results showed bilateral breast hyperplasia, abnormal seminal vesicle glands, rudimentary uterus, and underdeveloped right testis. Intraoperative examination revealed that the child had an ovary on the left and a testis on the right. The pathological results showed fibroadenomatoid changes in the breast. The patient had a karyotype of 46,XX. FISH results showed 46,XX.ish(DXZ1x2, SRYx0). Molecular testing showed that NR0B1, PHEX, CXORF21, GJB1, PQBP1, and COL4A5 genes are duplicated. There was a presence of SRY gene and absence of UYT gene. CONCLUSION DSD should be considered in patients with genital abnormality and male breast development. Ultrasound, sex hormone test and genetic testing should be performed to confirm the diagnosis of DSD, and molecular testing should be performed if necessary. Individualized treatment of DSD patient requires cooperation of multiple clinical disciplines.
Child, Preschool ; DNA Copy Number Variations ; DNA-Binding Proteins/genetics* ; Disorders of Sex Development/genetics* ; Female ; Genetic Testing ; Gonadal Steroid Hormones ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Sexual Development/genetics*