Objective:To investigate the clinical and genetic features of a Chinese pedigree with Progressive familial intrahepatic cholestasis (PFIC) and explore its genotype-phenotype correlation.Methods:A patient with PFIC diagnosed at Xinxiang Central Hospital in 2023 was selected as the study subject. The patient was subjected to abdominal magnetic resonance imaging (MRI) and painless gastroscopy. Peripheral blood samples were collected from the patient and his parents for the extraction of genomic DNA and trio-whole exome sequencing (trio-WES). Candidate variants were verified by Sanger sequencing. This study has been approved by the Medical Ethics Committee of Xinxiang Hospital (Ethics No. 2023-241).Results:MRI scan showed that the patient had significantly enlarged liver and spleen. WES revealed that he has harbored compound heterozygous variants of the ATP8B1 gene, including a c. 1710_1711insCCTC (p.A571Pfs*12) frameshifting variant in exon 16 and a c. 2989G>A (p.V997M) missense variant in exon 24, which were respectively inherited from his father and mother, and rated as pathogenic (PVS1+ PM2_Supporting+ PM3+ PP1) and likely pathogenic (PM2_Supporting+ PM3+ PP1) based on the guidelines from the American College of Medical Genetics and Genomics (ACMG). Conclusion:WES can clarify the genetic etiology of patients with speed and accuracy, and facilitate clinical decision-making. The detection of pathogenic variants has provided a basis for clinical diagnosis and enriched the mutational spectrum of the ATP8B1 gene.

OBJECTIVE: To investigate the clinical and genetic features of a Chinese pedigree with Progressive familial intrahepatic cholestasis (PFIC) and explore its genotype-phenotype correlation. METHODS: A patient with PFIC diagnosed at Xinxiang Central Hospital in 2023 was selected as the study subject. The patient was subjected to abdominal magnetic resonance imaging (MRI) and painless gastroscopy. Peripheral blood samples were collected from the patient and his parents for the extraction of genomic DNA and trio-whole exome sequencing (trio-WES). Candidate variants were verified by Sanger sequencing. This study has been approved by the Medical Ethics Committee of Xinxiang Hospital (Ethics No. 2023-241). RESULTS: MRI scan showed that the patient had significantly enlarged liver and spleen. WES revealed that he has harbored compound heterozygous variants of the ATP8B1 gene, including a c.1710_1711insCCTC (p.A571Pfs*12) frameshifting variant in exon 16 and a c.2989G>A (p.V997M) missense variant in exon 24, which were respectively inherited from his father and mother, and rated as pathogenic (PVS1+PM2_Supporting+PM3+PP1) and likely pathogenic (PM2_Supporting+PM3+PP1) based on the guidelines from the American College of Medical Genetics and Genomics (ACMG). CONCLUSION WES can clarify the genetic etiology of patients with speed and accuracy, and facilitate clinical decision-making. The detection of pathogenic variants has provided a basis for clinical diagnosis and enriched the mutational spectrum of the ATP8B1 gene.
Humans ; Male ; Pedigree ; Cholestasis, Intrahepatic/diagnostic imaging* ; Adenosine Triphosphatases/genetics* ; Female ; Adult ; Exome Sequencing ; Mutation ; East Asian People

Objective To evaluate the short-term efficacy of the VISULYZE software generated nomogram in assis-ting small incision lenticule extraction(SMILE)for the correction of myopia and astigmatism.Methods Non-randomized controlled trial.Patients who underwent SMILE surgery with the original nomogram,assisted by the same surgeon at the Laser Myopia Treatment Center of Xi'an NO.1 Hospital between February 2023 and January 2024,were included.A total of 52 patients(102 eyes)of myopic astigmatism with 3-month postoperative follow-up were collected.VISULYZE software was then used to generate a new nomogram.Subsequently,a total of 40 patients(70 eyes)with myopic with-the-rule astigmatism and a preoperative cylinder of ≤2.00 D,who underwent SMILE assisted by the new nomogram at the same center between August and November 2024,were enrolled.Among them,50 eyes had a target refraction of plano and were assigned to the experimental group.In addition,from the database of patients who underwent SMILE assisted by the origi-nal nomogram,42 patients(70 eyes)with myopic with-the-rule astigmatism and a cylinder of ≤2.00 D were screened,of which 51 eyes had a target refraction of plano,and these were assigned to the control group.The postoperative visual and refractive outcomes of both groups were compared at 3 months.Astigmatism results were analyzed using Alpins vector analysis.Results At 3 months postoperatively,among eyes with a target refraction of plano,50 eyes(98.0％)in the control group and all 50 eyes(100.0％)in the experimental group achieved an uncorrected distance visual acuity(UDVA)of ≥ 20/20.No eye in either group experienced a loss of more than one line in corrected distance visual acuity(CDVA)compared with the preoperative level.At 3 months postoperatively,63 eyes(90.0％)in the control group and 66 eyes(94.3％)in the experimental group had a spherical equivalent(SE)within-0.50 to 0.50 D.The postoperative cylinder was significantly lower in the experimental group than in the control group(P＜0.05).Vector analysis revealed that the ex-perimental group had smaller values for the difference vector,index of success,and absolute angle of error than the control group,with all differences being statistically significant(all P＜0.05).At 3 months postoperatively,43 eyes(61.4％)in the control group and 57 eyes(81.4％)in the experimental group had an angle of error within-5° to 5°.Conclusion The use of the VISULYZE software generated nomogram can optimize SMILE surgery design,offering good efficacy,safety,and predictability,and improving the precision of SMILE surgery for correcting myopia and astigmatism.

Objective To evaluate the short-term efficacy of the VISULYZE software generated nomogram in assis-ting small incision lenticule extraction(SMILE)for the correction of myopia and astigmatism.Methods Non-randomized controlled trial.Patients who underwent SMILE surgery with the original nomogram,assisted by the same surgeon at the Laser Myopia Treatment Center of Xi'an NO.1 Hospital between February 2023 and January 2024,were included.A total of 52 patients(102 eyes)of myopic astigmatism with 3-month postoperative follow-up were collected.VISULYZE software was then used to generate a new nomogram.Subsequently,a total of 40 patients(70 eyes)with myopic with-the-rule astigmatism and a preoperative cylinder of ≤2.00 D,who underwent SMILE assisted by the new nomogram at the same center between August and November 2024,were enrolled.Among them,50 eyes had a target refraction of plano and were assigned to the experimental group.In addition,from the database of patients who underwent SMILE assisted by the origi-nal nomogram,42 patients(70 eyes)with myopic with-the-rule astigmatism and a cylinder of ≤2.00 D were screened,of which 51 eyes had a target refraction of plano,and these were assigned to the control group.The postoperative visual and refractive outcomes of both groups were compared at 3 months.Astigmatism results were analyzed using Alpins vector analysis.Results At 3 months postoperatively,among eyes with a target refraction of plano,50 eyes(98.0％)in the control group and all 50 eyes(100.0％)in the experimental group achieved an uncorrected distance visual acuity(UDVA)of ≥ 20/20.No eye in either group experienced a loss of more than one line in corrected distance visual acuity(CDVA)compared with the preoperative level.At 3 months postoperatively,63 eyes(90.0％)in the control group and 66 eyes(94.3％)in the experimental group had a spherical equivalent(SE)within-0.50 to 0.50 D.The postoperative cylinder was significantly lower in the experimental group than in the control group(P＜0.05).Vector analysis revealed that the ex-perimental group had smaller values for the difference vector,index of success,and absolute angle of error than the control group,with all differences being statistically significant(all P＜0.05).At 3 months postoperatively,43 eyes(61.4％)in the control group and 57 eyes(81.4％)in the experimental group had an angle of error within-5° to 5°.Conclusion The use of the VISULYZE software generated nomogram can optimize SMILE surgery design,offering good efficacy,safety,and predictability,and improving the precision of SMILE surgery for correcting myopia and astigmatism.

Objective:To investigate the clinical and genetic features of a Chinese pedigree with Progressive familial intrahepatic cholestasis (PFIC) and explore its genotype-phenotype correlation.Methods:A patient with PFIC diagnosed at Xinxiang Central Hospital in 2023 was selected as the study subject. The patient was subjected to abdominal magnetic resonance imaging (MRI) and painless gastroscopy. Peripheral blood samples were collected from the patient and his parents for the extraction of genomic DNA and trio-whole exome sequencing (trio-WES). Candidate variants were verified by Sanger sequencing. This study has been approved by the Medical Ethics Committee of Xinxiang Hospital (Ethics No. 2023-241).Results:MRI scan showed that the patient had significantly enlarged liver and spleen. WES revealed that he has harbored compound heterozygous variants of the ATP8B1 gene, including a c. 1710_1711insCCTC (p.A571Pfs*12) frameshifting variant in exon 16 and a c. 2989G>A (p.V997M) missense variant in exon 24, which were respectively inherited from his father and mother, and rated as pathogenic (PVS1+ PM2_Supporting+ PM3+ PP1) and likely pathogenic (PM2_Supporting+ PM3+ PP1) based on the guidelines from the American College of Medical Genetics and Genomics (ACMG). Conclusion:WES can clarify the genetic etiology of patients with speed and accuracy, and facilitate clinical decision-making. The detection of pathogenic variants has provided a basis for clinical diagnosis and enriched the mutational spectrum of the ATP8B1 gene.

Objective To study the effect of lipopolysaccharide(LPS)-induced M1 polarization of macrophages on tumor growth.Methods Female Balb/c mice were randomly divided into control group and experimental group.Renca cells were used to establish subcutaneous tumor model.NaCl(100μl/mice,once every two days)or LPS(100μg/mice,once every two days)was injected to the tumor side when the tumor grew to 50mm3.The M1 polarization level of tumor-associated macrophages was detected by flow cytometry.Mouse tumor cell lines(ML-1,MC38,Renca)were divided into three groups:blank group(complete medium with 50％DMEM basal medium),control group(complete medium with 50％medium supernatant of cultured macrophages)and experimental group(complete medium with 50％medium supernatant of LPS pretreated cultured macrophages).The proliferation of tumor cells was detected by cell counting kit-8(CCK-8).The cell cycle and apoptosis of tumor cells were detected by flow cytometry.Results The proportion of tumor-associated macrophages M1 increased after LPS treatment(P<0.001),thus enhancing its anti-tumor function.LPS-induced M1 polarization of macrophages can significantly inhibit the proliferation of tumor cells(Renca:P=0.023,ML-1:P=0.045).LPS-induced M1 polarization of macrophages blocked G0/G1 phase(MC38:P=0.011,ML-1:P=0.022)or S phase(Renca:P=0.022)of tumor cell cycle,and then cell division was inhibited.LPS-induced M1 polarization of macrophages significantly induced apoptosis of tumor cells(Renca:P=0.04,ML-1:P=0.007).Conclusion LPS can play an anti-tumor role by inducing M1 polarization of macrophages.

Objective:To construct a recombinant herpes simplex virus 2 (HSV-2) expressing enhanced green fluorescent protein (EGFP) using clustered, regularly interspaced, short palindromic repeat/CRISPR-associated nuclease 9 (CRISPR/Cas9) technology.Methods:Four strategies for inserting exogenous EGFP gene into HSV-2 genome using CRISPR/Cas9 technology were designed: (1) conventional homology-directed repair: circular two homology arm donor-mediated gene knock-in; (2) linearized single homology arm donor-mediated gene knock-in; (3) homology-independent targeted integration; (4) conventional homology-directed repair-mediated by cell lines stably expressing Cas9 and sgRNA.Results:The recombinant virus HSV-2-EGFP was successfully constructed based on the second, the third and the fourth strategies. The second strategy was the most efficient, followed by the third and the fourth strategies. The purified recombinant virus could stably express green fluorescent protein in seven passages and shared similar growth characteristics in Vero cells to the parental virus.Conclusions:Linearized single homology arm donor could increase the efficiency of gene knock-in, and cell lines stably expressing Cas9 and sgRNA could increase the efficiency of gene knock-in mediated by homology-directed repair.

Objective:To investigate the effects of genomic location of a foreign gene in Shanghai-191 strain measles virus (MV) vector on gene expression and virus replication.Methods:The nucleotide sequence encoding S1 protein of SARS-CoV-2 was inserted at different positions in MV antigenome (the upstream of the N gene, between P and M genes, between H and L genes), and co-transfected into 293T cells with helper plasmids coding T7 RNA polymerase and N, P, and L proteins, respectively. The transfected cells were lysed and the supernatants were used to infected Vero cells to harvest recombinant viruses. S1 proteins expressed by the recombinant viruses were identified by RT-PCR, indirect immunofluorescence assay, Western blot and ELISA. Growth kinetics of the recombinant viruses were analyzed.Results:Recombinant viruses were failed to be rescued when the S1 protein-coding sequence was cloned into the upstream of N gene. Two recombinant viruses, MV-M-S1 and MV-L-S1, were successfully rescued when cloning the S1 protein-coding sequence into the intergenic region between P and M genes, or H and L genes, and could express S1 protein. MV-M-S1 expressed more S1 protein than MV-L-S1, but the titer of MV-M-S1 was lower.Conclusions:Inserting a foreign gene at different positions in the MV genome might have different effects on gene expression and virus replication. This study provided reference for the subsequent construction of MV vector.

Objective:To understand the basic situation of noise hazards in wood furniture manufacturing, stone processing and special equipment manufacturing industries in Shenzhen City, and to provide basis for preventing and controlling noise hazards.Methods:In August 2019, stratified random sampling ws used to select 21 wood funiture manafacturing, 15 stone processing and 24 special equipment manufactruing as survey object. Through the methods of occupational health field investigation, noise intensity measurement and occupational health examination data collection, the noise hazards of 60 enterprises in three industries were statistically analyzed.Results:There were 154, 44 and 113 noise detection points in wood furniture manufacturing, stone processing and special equipment manufacturing enterprises, and 101, 27 and 78 out of standard points respectively, and the exceeding standard rates were 65.6% (101/154) , 61.4% (27/44) and 69.0% (78/113) , respectively. There were 1670, 172 and 856 workers exposed to noise in the three industries. Among them, 1198, 134 and 703 people had taken occupational health examination. The physical examination rates were 71.7% (1198/1670) , 77.9% (134/172) and 82.1% (703/856) , respectively. The abnormal rates were 19.1% (229/1198) , 51.5% (69/134) and 37.8% (266/703) , respectively.Conclusion:High intensity noise working environment, too long noise exposure time, incorrect wearing of personal protective equipment and neglect of occupational health examination are important reasons for hearing loss of noise exposed personnel.

Objective:To understand the basic situation of noise hazards in wood furniture manufacturing, stone processing and special equipment manufacturing industries in Shenzhen City, and to provide basis for preventing and controlling noise hazards.Methods:In August 2019, stratified random sampling ws used to select 21 wood funiture manafacturing, 15 stone processing and 24 special equipment manufactruing as survey object. Through the methods of occupational health field investigation, noise intensity measurement and occupational health examination data collection, the noise hazards of 60 enterprises in three industries were statistically analyzed.Results:There were 154, 44 and 113 noise detection points in wood furniture manufacturing, stone processing and special equipment manufacturing enterprises, and 101, 27 and 78 out of standard points respectively, and the exceeding standard rates were 65.6% (101/154) , 61.4% (27/44) and 69.0% (78/113) , respectively. There were 1670, 172 and 856 workers exposed to noise in the three industries. Among them, 1198, 134 and 703 people had taken occupational health examination. The physical examination rates were 71.7% (1198/1670) , 77.9% (134/172) and 82.1% (703/856) , respectively. The abnormal rates were 19.1% (229/1198) , 51.5% (69/134) and 37.8% (266/703) , respectively.Conclusion:High intensity noise working environment, too long noise exposure time, incorrect wearing of personal protective equipment and neglect of occupational health examination are important reasons for hearing loss of noise exposed personnel.