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Objective To study the optimization measures for treating waterlogged collapsible loess stratum, it is necessary to investigate the deformation characteristics of tunnel structures after loess collapse by waterlogging. Method The foundation treatment for the waterlogged collapsible loess tunnels is introduced. Taking a shield metro tunnel crossing through waterlogged collapsible loess stratum as example, the method of three-axis mixing pile is adopted as the foundation treatment method. By establishing a numerical calculation model using MIDAS GTS software， the influence of waterlogged deformation of loess foundation on the vertical displacement of various metro tunnel structure parts and the internal force of lining structure is analyzed under different treatment depths and treatment widths. The treatment scheme is optimized based on the changing trends of tunnel parts vertical displacement and lining structure internal force, the construction economy and relevant requirements. Under conditions of three treatment widths and seven treatment depths, variation laws of vertical displacement, bending moment, and principal stress at the tunnel arch waist, vault, and invert are analyzed for the most suitable treatment recommendation. Result & Conclusion The greater the treatment depth, the smaller the vertical displacement of tunnel structure parts and the larger the lining structure internal force; the larger the treatment width, the smaller the vertical displacement and the internal force mentioned above. In this case study engineering, the optimal treatment depth is 7 m, and the optimal treatment width is 16 m.

Background Whole-course management refers to a patient-centered care model over the entire course of patients, aimed at patients' satisfaction with medical care, improving the whole course management of the disease. The whole-course management model has achieved significant success in various medical fields, such as the treatment of mental health disorders. However, its application in the general practitioner training has been rarely studied. Objective To investigate the practical effectiveness of the whole-course management teaching model in standardized training for general practitioners. Methods A total of 60 general practitioners of grades 2020, 2021 and 2022 who received national standardized training at Xiangya Second Hospital of Central South University. Additionally, 30 mentors specializing in total disease management were selected, with each mentor supervising two general practitioners, one from the research group and one from the control group, in order to minimize the impact of personal factors of mentors and ensure more objective research results. All levels of general practitioners of grades 2020, 2021 and 2022 were randomly divided into the research and control groups. The research group adopted whole-course management teaching mode while the control group followed the traditional lecture-based learning (LBL) model, with one team leader selected from each grade. After a nine-month training period (July 2022 to March 2023) , participants were evaluated on 6 items of diagnostic capability, treatment effect evaluation, patient management ability, teamwork ability, self-learning capability, and teaching ability. Simultaneously, a self-designed satisfaction questionnaire was also used to investigate patients under the care of these general practitioners. The average score was calculated based on ratings from three patients per general practitioner. Results Diagnostic capability, patient management ability, self-learning capability, and teaching ability scores of grade 2020 in the research group were high than those of the control group (P

The aim of this study was to investigate the antioxidant and anti-aging effects and underlying mechanisms of inactivated Bacillus sp. DU-106 in Caenorhabditis elegans. The antioxidant activity of inactivated Bacillus sp. DU-106 was evaluated by in vitro free radical scavenging assay. The effects of inactivated Bacillus sp. DU-106 at various concentrations on the lifespan, stress response and antioxidant capacity of C. elegans were observed. The results illustrated that inactivated Bacillus sp. DU-106 possessed strong free radical scavenging capacity in vitro, meanwhile, inactivated Bacillus sp. DU-106 effectively extended the lifespan of C. elegans and improved its movement and stress resistance. Different concentrations of inactivated Bacillus sp. DU-106 increased the levels of catalase (CAT), superoxide dismutase (SOD) and glutathione in a dose-dependent manner. Fluorescence microscopy showed that treatment with inactivated Bacillus sp. DU-106 significantly reduced lipofuscin levels in C. elegans. In conclusion, inactivated Bacillus sp. DU-106 can exert an anti-aging effect perhaps by reducing lipofuscin accumulation and increasing the levels of antioxidant enzymes in C. elegans. This study provides a powerful basis for the development of Bacillus for application in nutraceuticals.

Tumor-induced osteomalacia (TIO) is a rare paraneoplastic syndrome caused by tumors secreting fibroblast growth factor 23 (FGF23) that promotes urinary phosphorus excretion. Thus, TIO is typically characterized by phosphoruria, hypophosphatemia, and osteomalacia. Diagnosis and localization of the tumor is often difficult due to its small size, slow growth and concealed location. Due to the high expression of somatostatin receptors in pathogenic tumors, nuclear medicine functional imaging, particularly somatostatin receptor imaging, is used for diagnosis and localization of culprit tumors with high sensitivity and specificity. Here we retrospectively analyze 25 cases in which 68Ga-DOTATATE PET/CT successfully localized and diagnosed TIO culprit tumors. The clinical features, pathological results and image characteristics of 68Ga-DOTATATE PET/CT imaging were analyzed and compared with other imaging diagnostic techniques. It was confirmed that 68Ga-DOTATATE PET/CT imaging was the preferred imaging technique for successful diagnosis and localization of TIO pathogenic tumors.

Objective To investigate the laboratory and clinical characteristics of acute lymphoblastic leukemia (ALL) with e1a3 BCR-ABL fusion protein. Methods The clinical data, and results of bone marrow cytology, flow cytometry, karyotyping, fluorescence in situ hybridization (FISH) and real-time fluorescence quantitative PCR of 2 ALL cases with e1a3 BCR-ABL fusion protein admitted in our department were collected and analyzed. The related literature was reviewed. Results Both cases presented ALL phenotype. FISH test showed BCR-ABL positive, and RQ-PCR (p190, p210, p230) negative. Finally, the rare BCR-ABL fusion gene test was added and the 2 patients were diagnosed as rare e1a3 BCR-ABL positive ALL. One patient received TKI treatment and had achieved complete remission for more than 4 months. Another patient did not receive TKI and died in longer than 1 month after diagnosis. Conclusion Multi-technique joint detection for fusion gene is helpful for the detection and diagnosis of ALL with e1a3 BCR-ABL fusion protein, and TKI therapy is helpful to improve the prognosis.

Objective To clarify the pathogenic mutations and provide references for prevention and treatment strategies and prenatal diagnosis by gene detection in 3 pedigrees with autosomal dominant polycystic kidney disease (ADPKD). Methods The family history and clinical data of 3 ADPKD pedigrees who admitted to our department of urology during 2017 and 2020 were collected in this study. The peripheral blood samples of these family members were harvested for DNA extraction. Targeted amplification and high-throughput sequencing was used to screen suspected pathogenic mutations in probands, and then these obtained mutations were verified and analyzed in the probands and their family members by Sanger sequencing. The pathogenicity of these mutations was analyzed with bioinformation analysis and the evaluation criteria of the American Society of Medical Genetics. Results Ultrasound examination showed that all the probands from the 3 families had polycystic kidneys, at the clinical stages of G1, G3a, and G5 respectively. The deletion frameshift mutation c.5933delA (p.Asn1978fs), nonsense heterozygous mutation c.6871C>T (G2291X) and deletion frameshift mutation of c.894_897delCCCT (p.299Sfs*34) in polycystic kidney disease 1 (PKD1) gene were detected in pedigrees 1, 2 and 3, respectively. The above variants in the 3 families were in line with phenotype-genotype separation. Conclusion We identify the pathogenic variants in the 3 pedigrees of hereditary polycystic kidney disease, among which c.5933delA and c.894_897delCCCT are new mutations, and c.6871C>T is a known pathogenic mutation.

Objective: To explore the value of whole exome sequencing for the inferential analysis of recessive genetic disease carrier status for couples with a child died of Primary immunodeficiency (PID)., Methods: Clinical data was collected from four couples with a childbearing history of PID who had sought genetic counseling and undergone genetic testing at Henan Provincial People's Hospital from February 2017 to December 2021. Whole exome sequencing (WES) was performed on both partners of each couple, and candidate variants were validated by Sanger sequencing and fluorescent quantitative PCR. Prenatal diagnosis was conducted on fetuses of these couples after confirming the variants., Results: A total of six variants were detected in four genes including IL2RG, BTK, CYBB, and DUOX2. Among these, the c.1265G>A and c.3329G>A variants of the DUOX2 gene and the c.676C>T variant of the IL2RG gene were previously known as pathogenic variants. On the other hand, the Exon5_8del variant of the IL2RG gene, the c.184_185delAC variant of the BTK gene, and the c.472A>T variant of the CYBB gene were unreported previously. Based on the guidelines from the American College of Medical Genetics and Genomics, the IL2RG: Exon5_8del, BTK: c.184_185delAC and CYBB: c.472A>T variants were classified as likely pathogenic (PVS1+PM2_Supporting+PP4).Prenatal diagnosis was conducted for three couples during their subsequent pregnancies, and the results revealed that the fetuses had the wild-type genotypes at the c.184_185 position of the BTK gene, the c.472 position of the CYBB gene, and the c.676 position of the IL2RG gene. Follow-up examinations one year after birth has found no abnormality in the infants., Conclusion: WES is an important tool to infer and analyze the carrier status for couples who had given births to children died of PID and improve the positive detection rate.

Objective: To explore the clinical features and genetic etiology of two children with intellectual developmental disorder and microcephaly with pontine and cerebellar hypoplasia (MICPCH)., Methods: Two children with MICPCH who were presented at the Henan Provincial People's Hospital between April 2019 and December 2021 were selected as the study subjects. Clinical data of the two children were collected, along with peripheral venous blood samples of them and their parents, and amniotic fluid sample of the mother of child 1. Whole exome sequencing (WES), array-comparative genomic hybridization (aCGH) and real-time quantitative PCR (qPCR) were carried out for the children, their parents and the fetus. The pathogenicity of candidate variants were evaluated., Results: Child 1 was a 6-year-old girl featuring motor and language delay, whilst child 2 was a 4.5-year-old girl mainly featuring microcephaly and mental retardation. WES revealed that child 2 has harbored a 158.7 kb duplication in Xp11.4 (chrX: 41446160_41604854), which has encompassed exons 4~14 of the CASK gene. The same duplication was not found in either of her parents. aCGH revealed that child 1 has harbored a 29 kb deletion at Xp11.4 (chrX: 41637892_41666665), which encompassed exon 3 of the CASK gene. The same deletion was not found in either of her parents and the fetus. The above results were confirmed by qPCR assay. Above deletion and duplication were not found in the ExAC, 1000 Genomes and gnomAD databases. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), both variants were rated as likely pathogenic (PS2+PM2_Supporting)., Conclusion: The deletion of exon 3 and duplication of exons 4~14 of the CASK gene probably underlay the pathogenesis of MICPCH in these two children, respectively.

Objective: To analyze the clinical features and genetic variant in a patient with Usher syndrome., Methods: Whole exome sequencing was carried out for the patient. Suspected variants were validated by Sanger sequencing of her parents and fetus., Results: The proband was found to harbor compound heterozygous variants c.17_18insA (p.Tyr6Ter*) and c.4095_4096insA (p.Arg1366Lys fs*38) of the PCDH15 gene (NM_033056), which were respectively inherited from her father and mother. The same variants were not detected in 100 healthy controls. Based on the guidelines of the American Society of Medical Genetics and Genomics, both variants were predicted to be pathogenic (PVS1+PM2+PP4). By prenatal diagnosis, her fetus was found to carry the c.4095_4096insA variant. After birth, the child has passed neonatal hearing screening test, and no abnormal auditory and visual function was found after the first year., Conclusion: The compound heterozygous variants c.17_18insA (p.Tyr6Ter*) and c.4095_4096insA (p.Arg1366Lys fs*38) of the PCDH15 gene probably underlay the Usher syndrome is this proband.

Objectives: This study aims to investigate the effect of calbindin 1 on the proliferation and apoptosis of gingival epithelial cells affected by Porphyromonas gingivalis (P. gingivalis) in vitro ., Methods: A model of P. gingivalis infecting CA9-22 was established in vitro . At 24 h after infection, the expression of calbindin 1 (CALB1) was detected by real-time fluorescent quantitative polymerase chain reaction, Western blot, and immunofluorescence analyses. The expression of CALB1 was further inhibited by RNA interference. Cell proliferation was detected by BrdU analysis, and cell apoptosis was detected by caspase 3 activity. The expression of MDM2 and p53 was detected by Western blot analysis., Results: P. gingivalis infection upregulated the expression of CALB1 in CA9-22 cells with multiplicity-dependent manner. CALB1 promoted the proliferation of CA9-22 cells, increased the expression of MDM2, and inhibited the expression of p53. Inhibiting CALB1 expression did not affect the inhibitory effect of P. gingivalis infection on CA9-22 apoptosis., Conclusions: P. gingivalis infection can promote the proliferation of CA9-22 cells by increasing CALB1 expression. The related mechanism may be associated with MDM2-p53.

Objective: To explore the genetic basis for a Chinese pedigree affected with dyschromatosis symmetrica hereditaria (DSH)., Methods: PCR and Sanger sequencing were carried out for the proband, and suspected variant was validated by Sanger sequencing in the pedigree., Results: The proband was found to harbor a novel variant of c.1352delA (p.N451Mfs*13) of the ADAR (NM_001111) gene. The same variant was found in her affected mother and sister, but not in her unaffected father, uncle, and 100 healthy individual., Conclusion: The novel variant of the ADAR gene probably underlay the pathogenesis of DSH in this pedigree.

Objective: To explore the genetic basis for a pedigree affected with hereditary spastic paraplegia type 4 (HSP4)., Methods: Peripheral venous blood samples were taken from members of the four-generation pedigree and 50 healthy controls for the extraction of genomic DNA. Genes associated with peripheral neuropathy and hereditary spastic paraplegia were captured and subjected to targeted capture and next-generation sequencing. The results were confirmed by Sanger sequencing., Results: DNA sequencing suggested that the proband has carried a heterozygous c.1196C>G variant in exon 9 of the SPAST gene, which can cause substitution of serine by threonine at position 399 (p.Ser399Trp) and lead to change in the protein function. The same variant was also detected in other patients from the pedigree but not among unaffected individuals or the 50 healthy controls. Based on the ACMG 2015 guidelines, the variant was predicted to be possibly pathogenic., Conclusion: The c.1196C>G variant of the SPAST gene probably underlay the HSP4 in this pedigree.

Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose.

Objective: To detect variants of ARSA gene in a child featuring late infantile metachromatic leukodystrophy (MLD)., Methods: PCR and Sanger sequencing was carried out for the patient and her parents., Results: The patient had typical features of MLD including ARSA deficiency, regression of walking ability, and demyelination. Compound heterozygous variants of the ARSA gene, namely c.960G>A and c.244C>T, were detected in the patient, for which her mother and father were respectively heterozygous carriers. ARSA c.960G>A was known to be pathogenic, while ARSA c.244C>T was a novel variant. The same variants were not detected among 50 healthy controls., Conclusion: The compound heterozygous variants c.960G>A and c.244C>T of the ARSA gene probably underlie the MLD in this patient.

Objective: To explore the genetic basis for a pedigree affected with Bartter's syndrome (BS)., Methods: Panel-based next-generation sequencing (NGS) was carried out to detect mutation in BS-related genes SLC12A1, KCNJ1, BSND and CLCNKB. Sanger sequencing of MAGED2 gene and chromosomal microarray analysis (CMA) were also performed on the patient. Suspected mutation was validated in her family members., Results: No pathogenic mutation was detected by NGS, while a 0.152 Mb microdeletion at Xp11.21 (54 834 585-54 986 301) was found in the male fetus, which removed the entire coding region of the MAGED2 gene. His mother was a heterozygous carrier of the deletion. His father and sister did not carry the same deletion., Conclusion: The loss of the MAGED2 gene may underlie the BS in this pedigree.

Objective: To analyze the clinical features and genetic mutations in a patient with mucolipidosis type II α/β by using next generation sequencing., Methods: Clinical data of the patient was collected. Genomic DNA of the patient and her parents was extracted by a standard method. The patient was subjected to targeted sequencing using an Ion Ampliseq panel, which included genes related to mucolipidosis and mucopolysaccharidosis. Suspected mutations were verified by Sanger sequencing., Results: Compound heterozygous mutations, namely c.1284+1G>T and c.1090C>T (p.Arg364*), were detected in the patient, which were respectively inherited from her mother and father. No other disease-causing mutation was detected in the patient. GNPTAB c.1090C>T was known to be pathogenic, while GNPTAB c.1284+1G>T is a novel mutation. The same mutations were not detected among 50 healthy controls., Conclusion: The compound heterozygous mutations c.1284+1G>T and c.1090C>T (p.Arg364*) of GNPTAB gene probably account for the mucolipidosis type II α/β in the patient. NGS has a great value for the molecular diagnosis and typing of mucolipidosis.

Objective: To detect potential mutation of the WAS gene in a Chinese family affected with Wiskott-Aldrich syndrome., Methods: Peripheral blood samples were collected from the proband and his family members. All exons and flanking regions of the WAS gene were subjected to PCR amplification - Sanger sequencing as well as restriction endonuclease analysis. Plasma level of B-cell activating factor (BAFF) was also determined for all family members., Results: A hemizygous mutation (c.257G>A) of the WAS gene was identified in all patients from the family, for which the patient's mother was heterozygous. The same mutation was not found among healthy members of the family. Compared with unaffected members, all patients had a higher level of BAFF., Conclusion: The c.257G>A mutation of the WAS gene probably underlies the Wiskott-Aldrich syndrome in this family.

OBJECTIVE To detect potential mutations of the EXT1 and EXT2 genes in a pedigree affected with hereditary multiple exostosis (HME). METHODS For a four-generation family with 7 affected individuals from 17 family members,genomic DNA was extracted from peripheral venous blood samples. All exons of the EXT1 and EXT2 genes were screened for potential mutation by PCR and Sanger sequencing. RESULTS A novel heterozygous frameshift mutation c.1202delT (p.I401Tfs*2)was found in exon 4 of the EXT1 gene in the proband and the other 6 affected individuals. The same mutation was not detected among the healthy members from the family. The mutation has given rise a truncated EXT1 protein with loss of 345 amino acids. CONCLUSION A novel frameshift mutation of the EXT1 gene has been identified in a pedigree affected with HME, which has enriched the mutational spectrum of the EXT1 gene and may facilitate genetic counseling and prenatal diagnosis for the family.

Rutin-degrading enzymes (RDE) can degrade rutin into poorly water soluble compound, quercetin, and cause the bitter taste in tartary buckwheat. In the present study RDE from Yu 6-21 tartary buckwheat seeds was purified by ammonium sulphate precipitation, followed by hydrophobic interaction chromatography on Phenyl Sepharose CL-4B, ion exchange chromatography on CM-Cellulose and gel filtration chromatography on Sephadex G-150. Purified RDE showed single band with molecular weight of 66 kDa on SDS-PAGE. The optimum pH and temperature of RDE were 5.0 and 50 ℃ respectively. The Km was 0.27 mmol/L, and the Vmax was 39.68 U/mg. The RDE activity could be inhibited by Cu²⁺, Zn²⁺, Mn²⁺ and EDTA, and showed tolerance to 50% methanol (V/V). The N terminal sequence (TVSRSSFPDGFLFGL) was obtained by Edman degradation method and 15 internal peptide sequences were determined by MALDI-TOF-MS (matrix-assisted laser desorption ionization time of flight mass spectrometry). These results established the foundations for identification of the candidate gene of RDE via transcriptome data and further studying RDE biological function.

Viral protein(VP)16is an important tegument protein of the herpes virus. It is involved in early transcription activation of viral genes as well as virion assembly and release in host cells.VP16 of some herpes viruses have deubiquitinating protease activity and can help the virus counteract the host immune response. In this review, we explain the function and complex interactions between VP16 and other proteins based of the structural characteristics of VP16.This summary provides a reference for further research of maturation of the herpes virus as well as interactions between VP16 and other proteins.

Objective: To analyze the correlation between skeletal muscle mass and age among check-up adults., Methods: The study objects were those who aged 18-87 years old and came to a city hospital of Qiqihar for health examination from December, 2013 to September, 2014, excepted those with coronary heart disease, infectious disease, endocrine system disease, hypertension, anemia, cancer, liver disease, kidney disease and those carrying with heart pacemaker. A total of 1 836 respondents were finally enrolled into analysis. Appendicular Skeletal Muscle (ASM) was measured by a Body Composition Analyzer, and relative skeletal muscle index (RSMI) was calculated. The relationship among ASM, RSMI and age was assessed by linear regression analysis. The difference of height, weight, BMI, waist-hip-ratio (WHR), total muscle mass and percentage of body fat between genders were tested by t-test. The difference of ASM and total skeletal muscle mass between genders and among age groups was tested by multi-factor variance analysis. The difference of the muscle decline between genders was compared by Chi-square test., Results: The total muscle mass in males was (52.22 ± 6.65) kg, which was significantly higher than that in females ((38.05 ± 4.39) kg) (t=28.20, P<0.001). ASM in 18-29, 30-39, 40-49, 50-59, 60-69, 70-87 years was (24.64 ± 3.23), (24.00 ± 3.12), (24.35 ± 3.03), (23.33 ± 2.97), (22.54 ± 2.91) and (21.40 ± 3.36) kg (F=16.12, P<0.001) in males, respectively, and (16.48 ± 3.14), (16.72 ± 1.93), (16.75 ± 1.93), (16.84 ± 2.28), (16.52 ± 2.35) and(14.70 ± 2.37)kg (F=4.38, P=0.001) in females, respectively. ASM in males ((23.72 ± 3.16) kg) was higher than that in females ((16.65 ± 2.25) kg) (t=55.97, P<0.001). There was a negative correlation between age and ASM in males after 50 years old, the regression equation was y=28.31-0.09x (P<0.001). While a negative correlation between age and ASM in females occurred after 60 years old, the regression equation was y=27.69-0.18x (P<0.001). The prevalence of low ASM was 16.85% (124/736) in females, which was significantly higher than that in males (8.73%, 96/1 100) (χ(2)=27.57, P<0.001)., Conclusion: A negative correlation was found between age and ASM in males after 50 years old and in females after 60 years old. The prevalence of low RSMI in females was significantly higher than that in males.

Objective: To investigate the chemical constituents of the extract of Galium verum., Method: The compounds were isolated by chromatography and identified by spectral data., Result: The eleven compounds obtained were identified as (+)-pinoresinol 4,4'-O-bis-beta-D-glucopyranoside (1), epipinoresinol (2), (+) -medioresinol (3), isorhamnetin (4), isorhamnetin 3-O-alpha-L-rhamnopyranosyl-(1-6)-beta-D-glucopyranoside (5), diosmetin (6), diosmetin 7-O-beta-D-glucopyranoside (7), quercetin-3-O-beta-D-glucopyranoside (8), ursolic acid (9), ursolic aldehyde (10) and rubifolic acid (11)., Conclusion: Compounds 1-5 and 9-11 were isolated from this genus for the first time.

Objective: To understand the protective efficacy of China-made recombinant hepatitis B vaccine expressed by transgenic Chinese hamster ovary cell line among newborns., Methods: 2 969 newborns in seven townships in Zhengding County, Hebei Province, were vaccinated with 10 microgram x 3 doses of China-made recombinant hepatitis B vaccine expressed by transgenic Chinese hamster ovary cell line according to the 0 - 1 - 6 month schedule from 1 January 1997 to 31 August 1999. The newborns were to be vaccinated with the first dose within 24 hours after they were born. 1906 serum samples were selected in April 2000 to detect the hepatitis B infection markers, including HBsAg, HBsAb and HbcAb by RIA kits., Results: 2 783 of the 2 969 newborns (93.74%) were vaccinated with three doses, 2 833 of them (95.42%) were vaccinated with the first dose within 24 hours after they were born. The anti-HBs positive rate was 98.25% (S/N >/= 2.1) or 94.26% (S/N >/= 10.0), and the geometric mean titer (GMT) value of antibody was 77.64 within the first year after the whole course vaccination. Then the antibody level decreased gradually with the lapse of time. The HBsAb positive rate was 92.31% (S/N >/= 2.1) or 68.96% (S/N >/= 10.0), and GMT value was 22.86 within the third year after vaccination. The HBsAg positive rates remained less than 1%, the HBcAb positive rates and HBV infection rates remained 1% approximately 3% within 3 years after vaccination., Conclusion: The protective efficacy of China-made recombinant hepatitis B vaccine is satisfactory.