[Genetic and clinical analysis of a child with Shwachman-Diamond syndrome due to compound heterozygous variants of SBDS gene].

OBJECTIVE: To analyze the clinical features and genetic characteristics of a patient with Shwachman-Diamond syndrome (SDS) due to compound heterozygous variants of SBDS gene. METHODS: A female child with SDS who was admitted to the Children's Hospital Affiliated to Zhengzhou University in February 2022 was selected as the study subject. Clinical data of the child was collected. Peripheral blood samples of the child and her elder sister and parents were collected and subjected to whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing. RESULTS: The child, a 1-year-and-1-month-old girl, had mainly manifested with diarrhea, hematochezia, growth retardation and malnutrition, along with increased transaminases and decreased neutrophils and hemoglobin. Anteroposterior X-ray of her left wrist indicated significantly delayed bone age. Colonoscopy revealed that her colorectal mucosa was erosive with oily food residues attached to the intestinal lumen. Genetic testing revealed that she has harbored c.258+2T>C and c.100A>G compound heterozygous variants of the SBDS gene. The c.258+2T>C variant has derived from her father and known to be pathogenic, whilst the other has derived from her mother. Based on the guidelines from the American College of Medical Genetics and Genomics, the c.100A>G variant was classified as likely pathogenic (PM1+PM2_Supporting+PM3+PM5+PP3). CONCLUSION: The compound heterozygous variants of c.258+2T>C and c.100A>G probably underlay the SDS in this child. For children with refractory diarrhea, liver damage and growth retardation, SDS should be suspected, and genetic testing can facilitate the diagnosis and treatment.

[Optimization Design of Interbody Fusion Cage withDifferent Bone Densities Printed in 3D].

Based on the original data of cervical spine, the models of cervical C6 and C7 segments were reconstructed through medical image processing and reverse modeling operations, then the models were assembled to obtain the basic data of interbody fusion cage. According to the basic data, the structures of rectangular porous, gradient porous and octahedral porous interbody fusion cages were established respectively. Maximum force on the adult male neck was applied to the fusion device, and the stress, strain and maximum deformation of the fusion device were solved by finite element analysis. The elastic modulus decrease of the design, and the rectangular porosity structures with different porosity were analyzed and optimized. The results showed that the elastic modulus of the fusion cage with three structures decreased in varying degrees, and the porosity of the interbody fusion cage with rectangular structure was about 60%, which was the most decreased elastic modulus.

A diabetes prediction model based on Boruta feature selection and ensemble learning.

BACKGROUND AND OBJECTIVE: As a common chronic disease, diabetes is called the "second killer" among modern diseases. Currently, there is no medical cure for diabetes. We can only rely on medication for auxiliary treatment. However, many diabetic patients still die each year. In addition, a considerable number of people do not pay attention to their physical health or opt out of treatment due to lack of money, which eventually leads to various complications. Therefore, diagnosing diabetes at an early stage and intervening early is necessary; thus, developing an early detection method for diabetes is essential. METHODS: In this study, a diabetes prediction model based on Boruta feature selection and ensemble learning is proposed. The model contains the use of Boruta feature selection, the extraction of salient features from datasets, the use of the K-Means++ algorithm for unsupervised clustering of data and stacking of an ensemble learning method for classification. It has been validated on a diabetes dataset. RESULTS: The experiments were performed on the PIMA Indian diabetes dataset. The model was evaluated by accuracy, precision and F1 index. The obtained results show that the accuracy rate of the model reaches 98% and achieves good results. CONCLUSION: Compared with other diabetes prediction models, this model achieved better results, and the obtained results indicate that this model is superior to other models in diabetes prediction and has better performance.

Pathogenicity of Congenital Adrenal Hyperplasia Induced by the p.P377L Mutation of CYP11B1.

CYP11B1 encodes an 11ß-hydroxylase that is involved in the catalysis of adrenal glucocorticoids and the production of cortisol. Mutations in CYP11B1 can result in congenital adrenal hyperplasia. We discovered a proband with a CYP11B1 gene mutation. Gene sequencing revealed a homozygous missense mutation of c.1130C > T in the 7th exon of the CYP11B1 gene that resulted in the change from Pro377 to leucine in the encoded protein. Based on the proband's clinical symptoms and the prognosis according to the database, this mutation may be harmful. However, the pathogenicity has not yet been reported. Thus, we created an expression vector for the mutation in vitro, transfected cells, observed the changes in gene expression, and determined its pathogenicity. To determine the pathogenicity of the CYP11B1 p.P377L mutation site through in vitro verification. The eukaryotic expression vector of the CYP11B1 mutation site was constructed in vitro, and the success of the construct was confirmed by sequencing. Fluorescence microscopy was used to determine the transfection effectiveness, GFP fluorescent tag labeling was used to detect changes in protein localization, and qRT‒PCR and Western blotting were used to detect CYP11B1 mRNA and protein expression. Sequencing revealed that the proband harbored a homozygous missense mutation of CYP11B1 (p.P377L). The expression of the protein decreased but the localization did not change when cells were transfected with the CYP11B1 mutation vector compared to the wild-type vector. The p.P377L mutation of CYP11B1 could affect protein expression and enzymatic activity and may be pathogenic.

Mercator: a pipeline for multi-method, unsupervised visualization and distance generation.

SUMMARY: Unsupervised machine learning provides tools for researchers to uncover latent patterns in large-scale data, based on calculated distances between observations. Methods to visualize high-dimensional data based on these distances can elucidate subtypes and interactions within multi-dimensional and high-throughput data. However, researchers can select from a vast number of distance metrics and visualizations, each with their own strengths and weaknesses. The Mercator R package facilitates selection of a biologically meaningful distance from 10 metrics, together appropriate for binary, categorical and continuous data, and visualization with 5 standard and high-dimensional graphics tools. Mercator provides a user-friendly pipeline for informaticians or biologists to perform unsupervised analyses, from exploratory pattern recognition to production of publication-quality graphics. AVAILABILITYAND IMPLEMENTATION: Mercator is freely available at the Comprehensive R Archive Network (https://cran.r-project.org/web/packages/Mercator/index.html).

Distinguish the Value of the Benign Nevus and Melanomas Using Machine Learning: A Meta-Analysis and Systematic Review.

Background: Melanomas, the most common human malignancy, are primarily diagnosed visually, beginning with an initial clinical screening and followed potentially by dermoscopic analysis, a biopsy, and histopathological examination. We aimed to systematically review the performance and quality of machine learning-based methods in distinguishing melanoma and benign nevus in the relevant literature. Method: Four databases (Web of Science, PubMed, Embase, and the Cochrane library) were searched to retrieve the relevant studies published until March 26, 2022. The Predictive model Deviation Risk Assessment tool (PROBAST) was used to assess the deviation risk of opposing law. Result: This systematic review included thirty researches with 114007 subjects and 71 machine learning models. The convolutional neural network was the main machine learning method. The pooled sensitivity was 85% (95% CI 82-87%), the specificity was 86% (82-88%), and the C-index was 0.87 (0.84-0.90). Conclusion: The findings of our study showed that ML algorithms had high sensitivity and specificity for distinguishing between melanoma and benign nevi. This suggests that state-of-the-art ML-based algorithms for distinguishing melanoma from benign nevi may be ready for clinical use. However, a large proportion of the earlier published studies had methodological flaws, such as lack of external validation and lack of clinician comparisons. The results of these studies should be interpreted with caution.

Serum pharmacochemistry and pharmacokinetics of major components after oral administration of Luhong recipe in rats using ultra-performance liquid chromatography-tandem mass spectrometry.

Luhong recipe (LHR) is has been used as an empirical prescription for treating chronic heart failure for long, with safety, reliability, and significant efficacy. However, its pharmacokinetics has not yet been studied. This study aims to establish a ultra performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous analysis of epimedin A, epimedin B, epimedin C, icariin, psoralen, and isopsoralen in rat plasma and apply it to the pharmacokinetic study of LHR after oral administration. These six analytes were ionized using positive electrospray ionization (ESI+ ). The MS/MS transitions used for monitoring are successively converted to m/z 839.3 → 369.1, m/z 809.2 → 369.1, m/z 823.3 → 369.1, m/z 677.2 → 205.2, m/z 187.1 → 115.2, and m/z 230 → 120.9. Linearity, precision, accuracy, stability, matrix effect, and recovery of the established method were within the acceptable range. The method was suitable for the determination of six analytes after oral administration of LHR. The pharmacokinetic results showed that the time to reach the peak concentration (Tmax ) was from 0.17 to 13.5 h, the peak concentration (Cmax ) was 109.23-980 ng/mL, the area under the concentration-time curve (AUC[0 - t] ) was 65.48-8846.08 ng·h/mL, and the apparent distribution volume (Vd) was 24,772-896,132 mL/kg. These results provided a meaningful basis for formulating the clinical dose regimen of LHR.

Breakpoints Identification of a Balanced Complex Chromosome Rearrangement Case: 46,XX, t(6;15;10;9)(q13;q15;p11.2;q34.3) ins(9;8)(q22.33;q21.1q21.3).

BACKGROUND: Balanced complex chromosome rearrangement (CCR) carriers are phenotypically normal but at high risk of reproductive failure, recurrent miscarriages, and affected offspring, so that cytogenetic characterizations of CCR carriers are crucial. METHODS: We report a case of CCR: 46,XX, t(6;15;10;9)(q13;q15;p11.2;q34.3) ins(9;8)(q22.33;q21.1q21.3). The peripheral blood was collected for karyotyping, single nucleotide polymorphism array (SNP-array) analysis, and whole genome mate-pair sequencing. RESULTS: The patient's karyotype is detected and identified as 46,XX, t(6;15;10;9)(q13;q15;p11.2;q34.3) ins(9;8) (q22.33;q21.1q21.3), with no significant duplication and deletion found by SNP-array analysis. There are 16 break-points among chromosomes 6, 8, 9, 10, and 15 identified by whole genome sequencing. CONCLUSIONS: With a variety of detection techniques, we can deeply study the genetic characteristics of CCRs, thus providing a basis for genetic counseling and choice of fertility.

Ang-(1-7) protects skeletal muscle function in aged mice.

BACKGROUND: The angiotensin-converting enzyme 2 (ACE2)/angiotensin 1-7 (Ang-(1-7)) axis has been shown to protect against the age-associated decline in skeletal muscle function. Here, we investigated the protective effects of ACE2 in mitigating the age-associated decline of skeletal muscle function and to identify the potential underlying molecular mechanisms. METHODS: We measured the expression levels of Ang-(1-7) in C57BL/6J mice of different ages and correlated these levels with measures of skeletal muscle function. We also investigated the expression of myocyte enhancer factor 2 A (MEF2A) in ACE2 knockout (ACE2KO) mice and its relationship with muscle function. We then treated aged ACE2KO mice for four weeks with Ang-(1-7) and characterized the levels of MEF2A and skeletal muscle function before and after treatment. We assessed the impact of Ang-(1-7) on the growth and differentiation of C2C12 cells in vitro and assessed changes in expression of the glucose transporter type 4 (Glut4). RESULTS: Aged mice showed reduced skeletal muscle function and levels of Ang-(1-7) expression in comparison to young and middle-aged mice. In ACE2KO mice, skeletal muscle function and MEF2A protein expression were significantly lower than in age-matched wild-type (WT) mice. After one month of Ang-(1-7) treatment, skeletal muscle function in the aged ACE2KO mice improved, while MEF2A protein expression was similar to that in the untreated group. In C2C12 cells, Ang-(1-7) was shown to promote along with the upregulated expression of Glut4. CONCLUSIONS: The ACE2/ Ang-(1-7) axis has a protective function in skeletal muscle and administration of exogenous Ang-(1-7) can delay the age-related decline in the function of skeletal muscle.

Effects of different nutritional conditions on accumulation and distribution of Cr in Coix lacryma-jobi L. in Cr6+-contaminated constructed wetland.

In this research, micro Coix lacryma-jobi L. vertical flow constructed wetlands (VFCWs) were set up using domestic sewage (DWS) and 1/2 Hoagland nutrient solution (HNS) as VFCWs water sources. 0, 20 mg L-1 and 40 mg L-1 of Cr6+ (in the form of K2Cr7O2) were added into the water sources separately in order to study the response of Coix lacryma-jobi L. under Cr6+ stress. The results showed that the inhibition rates of Cr6 + on plant height, stem diameter, shoot and root dry weight treated with HNS were 2.88~10.16%, 5.12~11.86%, 3.53~6.51% and 2.89~6.34% higher than those in DWS treatment. SEM analysis showed that the nuclear bilayer membrane was slightly damaged, the chromatin decreased and the number of mitochondrial cristae decreased when treated with 20 mg L-1 of Cr6+, however, organelle damage was more severe under 40 mg L-1 of Cr6+exposure. The X-ray energy spectrum analysis results indicated that the accumulation of chromium in epidermis and endodermis were higher than those in stele. The contents of total Cr in roots, stems and leaves treated with HNS were higher than those of DWS treatment. The highest content of Cr was observed in cell wall (32.12-188.1 mg kg-1), followed by vacuole (5.0-38.14 mg kg-1). The contents of Cr in each subcellular component in roots, stems, and leaves treated with HNS were higher than those of DWS, except for organelle components in the 14th week. DWS was used as water influent, the contents of easily migrated combined Cr (ETM) in roots, stems and leaves were significantly lower than those in HNS treatment. Improving the nutritional conditions of constructed wetlands might be beneficial to the improvement of their ability to purify chrome-containing waste water.

[Clinical characterization and genetic analysis of a newborn with chromosome 8q21.11 deletion syndrome].

OBJECTIVE: To explore the genetic etiology for a newborn with corneal opacity. METHODS: The neonate and her parents were subjected to routine G-banding chromosomal karyotyping analysis. Copy number variation (CNV) was analyzed with low-coverage whole-genome sequencing (WGS) and single nucleotide polymorphism microarray (SNP array). RESULTS: No karyotypic abnormality was found in the newborn and her parents. Low-coverage WGS has identified a de novo 5.5 Mb microdeletion at chromosome 8q21.11-q21.13 in the neonate, which encompassed the ZFHX4 and PEX2 genes. The result was confirmed by SNP array-based CNV analysis. CONCLUSION: The newborn was diagnosed with chromosome 8q21.11 deletion syndrome. ZFHX4 may be one of the key genes underlying this syndrome.

[Clinical features and genetic analysis of a child with late-onset immune dysregulation, polyendocrinopathy, enteropathy, X-Linked syndrome].

OBJECTIVE: To report on the clinical features and result of genetic testing for a child featuring immune dysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX) syndrome. METHODS: Clinical records, genetic testing, laboratory investigation and treatment of the child were summarized in addition with a comprehensive review of the literature. RESULTS: The 3-year-old boy was administered due to intractable diarrhea, recurrent infections, liver dysfunction and failure to thrive, though no diabetes or skin disorder was observed. Laboratory testing showed elevated liver enzymes and total IgE, decreased albumin and electrolyte imbalance. Gastrointestinal endoscopy revealed erosion and granules in the duodenum, and edema in the terminal ileum and colon. Biopsies showed villous atrophy in the duodenum and terminal ileum. Genetic testing revealed that the patient has carried a missense c.1087A>G (p.I363V) variant in the exon 10 of the FOXP3 gene. He was treated with enteral and parenteral nutrition, anti infection and Sirolimus, and was waiting for hemopoietic stem cell transplantation. CONCLUSION: Although IPEX syndrome usually occur during infancy, it should not be ruled out solely based on the age, and its presentation can be variable. For male children with refractory diarrhea, autoimmune disorder and growth retardation, the diagnosis should be suspected and confirmed by genetic testing.

CytoGPS: a web-enabled karyotype analysis tool for cytogenetics.

SUMMARY: Karyotype data are the most common form of genetic data that is regularly used clinically. They are collected as part of the standard of care in many diseases, particularly in pediatric and cancer medicine contexts. Karyotypes are represented in a unique text-based format, with a syntax defined by the International System for human Cytogenetic Nomenclature (ISCN). While human-readable, ISCN is not intrinsically machine-readable. This limitation has prevented the full use of complex karyotype data in discovery science use cases. To enhance the utility and value of karyotype data, we developed a tool named CytoGPS. CytoGPS first parses ISCN karyotypes into a machine-readable format. It then converts the ISCN karyotype into a binary Loss-Gain-Fusion (LGF) model, which represents all cytogenetic abnormalities as combinations of loss, gain, or fusion events, in a format that is analyzable using modern computational methods. Such data is then made available for comprehensive 'downstream' analyses that previously were not feasible. AVAILABILITY AND IMPLEMENTATION: Freely available at http://cytogps.org.

Different lipid profiles, insulin sensitivity, and insulin resistance among Han, Uygur, and Kazak men with normal glucose tolerance in Xinjiang, China.

BACKGROUND: This study aimed to determine the differences in clinical parameters among Han, Uygur, and Kazak men with normal glucose tolerance. METHODS: Participants' data from the China National Diabetes and Metabolic Disorders Study pertaining to Han, Uygur, and Kazak men from the Xinjiang province were used (n = 930). Pearson's correlation was used to examine the relationship between HOMA-IR, Matsuda Index, and clinical characteristics. RESULTS: HOMA-IR of Han men was significantly higher than in Uygurs and Kazaks (P < 0.001). The Matsuda Index of Kazaks was significantly higher than that of Hans and Uygurs (P < 0.001). While Kazaks had the highest BMI, WC, SBP, and DBP; they also had the highest HDL-C and lowest TG (P < 0.001). TG of Uygurs was significantly higher than that of Hans and Kazaks (P < 0.001). In Hans and Kazaks, the TG/HDL-C ratio increased with HOMA-IR quartiles; there was no association in Uygurs. In Hans and Kazaks, the TG/HDL-C ratio decreased with Matsuda index quartiles; there was no association in Uygurs. Multivariate linear regression showed that HOMA-IR was independently associated with ethnicity, BMI and TG/HDL-C ratio (P < 0.01), while Matsuda index was independently associated with ethnicity, BMI, LDL-C levels (P < 0.001) and TG/HDL-C ratio (P < 0.001). CONCLUSIONS: In conclusion, Han, Uygur, and Kazak men had different lipid profiles, BMI, and WC. Han men had the highest insulin resistance while Kazak men had the highest insulin sensitivity.

The relationship between the biovars and the antimicrobial resistance of Ureaplasma urealyticum in female patients with urogenital infections.

BACKGROUND: Ureaplasma urealyticum is considered as one of the main pathogens found in women with urogenital infection. This study aimed to investigate the relationship between the biovars, serovars, and their antimicrobial resistance against antibiotics in female patients with urogenital infection. METHODS: Two hundred and forty-six cervical secretion samples (125 female outpatients as the patient group, 121 healthy female subjects as the control group) were first collected and analyzed for U. urealyticum using the Mycoplasma Identification and Antimicrobial Susceptibility Testing; then polymerase chain reaction (PCR) was carried out to identify the biovars and serovars of U. urealyticum-positive samples. RESULTS: The prevalence of U. urealyticum in the patient group (57. 60%) was higher than that in the control group (24.79%, P<.01). The main biovar was biovar 1, and the main serovars were 1 (S1), 3 (S3), 6 (S6) in biovar 1. Mixed infection was observed in biovar 2. According to the results of Antimicrobial Susceptibility Testing in the patient group, biovar 1 shows more resistance to minocycline, doxycycline, and azithromycin than biovar 2 (P<.05). Serovars S1, S3 and S6 have the highest resistant rate to ofloxacin (84.38%), roxithromycin (84.62%), and azithromycin (90.90%), respectively. CONCLUSIONS: A high prevalence of U. urealyticum was observed in female patients with urogenital infections. And the biovar 1 and the serovars 1, 3, 6 were the main types of pathogens.

Chromium removal capability and photosynthetic characteristics of Cyperus alternifolius and Coix lacryma-jobi L. in vertical flow constructed wetland treated with hexavalent chromium bearing domestic sewage.

In this study, the chromium removal capability and photosynthetic capacity response of plants were investigated in vertical flow wetland microcosms (VFWM) treated with Cr(VI) bearing domestic sewage. Two plants, Cyperus alternifolius (C. alternifolius) and Coix lacryma-jobi L. (C. lacryma-jobi L.) grown in the VFWM enhanced the purification of Cr(VI) enriched domestic sewage. Cr concentration in the effluent fell below detection limit (<0.03 mg L-1), except for the C. alternifolius wetland treated with 40 mg L-1 Cr(VI). The biomasses of both plants species were increased at 10 and 20 mg L-1 Cr(VI) exposure but inhibited at 40 mg L-1 Cr(VI). The photosynthetic capacities of both plants were not affected at 10-40 mg L-1 Cr(VI) exposure during the days 20-60. However, they were inhibited significantly (P < 0.05) at 40 mg L-1 Cr(VI) exposure during days 80-100. These results demonstrated that a VFWM with C. alternifolius and/or C. lacryma-jobi L. was capable of maintaining its efficiency and recovering its vegetation. VFWM with C. alternifolius and/or C. lacryma-jobi L. was promising for purifying wastewater which contains low to medium concentrations of Cr(VI) (<20 mg L-1).

[Combined G-banded karyotyping and multiplex ligation-dependent probe amplification for the detection of chromosomal abnormalities in fetuses with congenital heart defects].

OBJECTIVE: To assess the value of G-banded karyotyping in combination with multiplex ligation-dependent probe amplification (MLPA) as a tool for the detection of chromosomal abnormalities in fetuses with congenital heart defects. METHODS: The combined method was used to analyze 104 fetuses with heart malformations identified by ultrasonography. Abnormal findings were confirmed with chromosomal microarray analysis (CMA). RESULTS: Nineteen (18%) fetuses were found to harbor chromosomal aberrations by G-banded karyotyping and MLPA. For 93 cases, CMA has detected abnormalities in 14 cases including 10 pathogenic copy number variations (CNVs) and 4 CNVs of uncertain significance (VOUS). MLPA was able to detect all of the pathogenic CNVs and 1 VOUS CNV. CONCLUSION: Combined use of G-banded karyotyping and MLPA is a rapid, low-cost and effective method to detect chromosomal abnormalities in fetuses with various heart malformations.

Study on the Process Characteristics Based on Joule Heat of Sliding-Pressure Additive Manufacturing.

This study developed an experimental system based on Joule heat of sliding-pressure additive manufacturing (SP-JHAM), and Joule heat was used for the first time to accomplish high-quality single-layer printing. The roller wire substrate is short-circuited, and Joule heat is generated to melt the wire when the current passes through. Through the self-lapping experimental platform, single-factor experiments were designed to study the effects of power supply current, electrode pressure, contact length on the surface morphology and cross-section geometric characteristics of the single-pass printing layer. Through the Taguchi method, the effect of various factors was analyzed, the optimal process parameters were obtained, and the quality was detected. The results show that with the current increase, the aspect ratio and dilution rate of a printing layer increase within a given range of process parameters. In addition, with the increase in pressure and contact length, the aspect ratio and dilution ratio decrease. Pressure has the greatest effect on the aspect ratio and dilution ratio, followed by current and contact length. When a current of 260 A, a pressure of 0.60 N and a contact length of 1.3 mm are applied, a single track with a good appearance, whose surface roughness Ra is 3.896 µm, can be printed. Additionally, the wire and the substrate are completely metallurgically bonded with this condition. There are also no defects such as air holes and cracks. This study verified the feasibility of SP-JHAM as a new additive manufacturing strategy with high quality and low cost, and provided a reference for developing additive manufacturing technology based on Joule heat.

Wolfram syndrome type 1: a case series.

BACKGROUND: Wolfram syndrome (WS) is a rare autosomal recessive multisystem neurodegenerative disease characterized by non-autoimmune insulin-dependent diabetes mellitus, optic atrophy, sensorineural deafness, and diabetes as the main features. Owing to clinical phenotypic heterogeneity, the misdiagnosis rate is high. However, early accurate diagnosis and comprehensive management are key to improving quality of life and prolonging life. RESULTS: Eleven patients from seven WS pedigrees with 10 mutation sites (c.1314_1317delCTTT, c.C529T, c.C529A, c.G2105A, c.C1885T, c.1859_1860del, c.G2020A, c.C529A, c.G2105A, and c.G1393C) in the WFS1 gene were included. We conducted further expert department analysis to clarify the diagnosis and analyze the correlation between genes and phenotypes. CONCLUSIONS: The genotypes of these patients were closely associated with their phenotypes. The clinical data of the patients were analyzed to provide a basis for the diagnosis and clinical management of the disease.

Integrated flow cytometric and proteomics analyses reveal the regulatory network underlying sugarcane protoplast responses to fusion.

BACKGROUND: Somatic cell fusion is a process that transfers cytoplasmic and nuclear genes to create new germplasm resources. But our limited understanding of the physiological and molecular mechanisms that shape protoplast responses to fusion. METHOD: We employed flow cytometry, cytology, proteomics, and gene expression analysis to examine the sugarcane (Saccharum spp.) protoplast fusion. RESULTS: Flow cytometry analysis revealed the fusion rate of protoplasts was 1.95%, the FSC value and SSC of heterozygous cells was 1.17-1.47 times higher than that of protoplasts. The protoplasts viability decreased and the MDA increased after fusion. During fusion, the cell membranes were perforated to different degrees, nuclear activity was weakened, while microtubules depolymerized and formed several short rod like structures in the protoplasts. The most abundant proteins during fusion were mainly involved in RNA processing and modification, cell cycle control, cell division, chromosome partition, nuclear structure, extracellular structures, and nucleotide transport and metabolism. Moreover, the expression of key regeneration genes, such as WUS, GAUT, CESA, PSK, Aux/IAA, Cdc2, Cyclin D3, Cyclin A, and Cyclin B, was significantly altered following fusion. PURPOSE AND SIGNIFICANCE: Overall, our findings provide a theoretical basis that increases our knowledge of the mechanisms underlying protoplast fusion.