Objective:To evaluate the role of whole exome sequencing（WES）in the etiological diagnosis and precision medicine management of patients with urolithiasis.Methods:We conducted a retrospective review of 21 patients with urolithiasis and pathogenic gene mutations identified by WES at The Second Affiliated Hospital of Zhengzhou University between April 2019 and March 2025. The cohort included 13 males and 8 females，with a mean age of（18.9 ± 11.1）years；18 patients were under 25 years old. Clinical presentations included nephrocalcinosis（8 patients）and urinary tract calculi（13 patients），with five patients exhibiting extra-renal manifestations such as renal tubular acidosis and hyperaldosteronism. Stone composition analysis identified calcium oxalate（16 patients），cystine（4 patients），and carbonate apatite（1 patient）. Metabolic abnormalities were prevalent，including hypocitraturia（11 patients），hyperoxaluria（8 patients），and hypercalciuria（7 patients），with eight patients presenting two or more concurrent disorders. All patients underwent WES and comprehensive metabolic evaluation. Sequencing was performed on an Illumina Hiseq4000 platform，achieving a mean depth of > 100× and coverage of > 98% in target regions. Variants were classified according to the American College of Medical Genetics and Genomics（ACMG）guidelines.Results:WES identified 12 distinct genes across autosomal recessive（9 genes： AGXT， GRHPR， ATP6V1B1， SLC12A1， KCNJ1， SLC3A1， SLC7A9， SLC34A3， WFS1），autosomal dominant（2 genes： CASR， ADCY10），and X-linked recessive（1 gene： CLCN5）inheritance patterns. Genotype-phenotype correlations revealed mutations associated with primary hyperoxaluria（8 patients），hypercalciuria（7 patients），and renal malformation due to a WFS1 mutation（1 patient）. A positive genetic diagnosis was achieved in 100% of patients with either urinary oxalate > 1 000 μmol/24 h or cystine stones. 8 patients received a diagnosis of hereditary stone disease at their first presentation（non-delayed group），while 13 experienced a mean diagnostic delay of（9.6 ± 3.9）years. The delayed diagnosis group had a significantly older age at initial stone onset［（10.2 ± 5.3）years vs.（6.8 ± 3.1）years， P = 0.03］and a higher incidence of impaired renal function（6 patients vs. 1 patient， P = 0.04）. Analysis of diagnostic delay by gene subgroup showed delays in 2/4 patients with cystinuria［ SLC3A1/ SLC7A9；（8.2 ± 3.5）years］，5/8 with primary hyperoxaluria［ AGXT/ GRHPR；（10.5 ± 4.1）years］，5/7 with hypercalciuria-related genes［ CASR/ ADCY10/ SLC12A1/ KCNJ1/ SLC34A3；（9.8 ± 3.8）years］，and 1/2 with other genes［ ATP6V1B1/ WFS1/ CLCN5；（7.6 ± 2.2）years］. Among 32 mutation sites detected，21 were classified as pathogenic/likely pathogenic and 11 as variants of uncertain significance. Four novel mutations were identified： ATP6V1B1（presenting with renal tubular acidosis，nephrocalcinosis，and hypocitraturia）， WFS1（presenting with renal malrotation，hydronephrosis，and stones without metabolic abnormalities）， SLC12A1（presenting with Bartter syndrome type 1，chronic renal insufficiency，hypercalciuria，hypocitraturia，alkalosis，and hyperaldosteronism），and SLC3A1（presenting with bilateral renal stones and cystinuria）. Conclusions:WES is crucial in identifying the underlying etiology of urolithiasis and can guide targeted treatment. We recommend early WES for patients with an initial stone presentation before age 25，those with nephrocalcinosis，or those with abnormal metabolic workups to facilitate precise diagnosis and preventive care.

Objective:To evaluate the role of whole exome sequencing（WES）in the etiological diagnosis and precision medicine management of patients with urolithiasis.Methods:We conducted a retrospective review of 21 patients with urolithiasis and pathogenic gene mutations identified by WES at The Second Affiliated Hospital of Zhengzhou University between April 2019 and March 2025. The cohort included 13 males and 8 females，with a mean age of（18.9 ± 11.1）years；18 patients were under 25 years old. Clinical presentations included nephrocalcinosis（8 patients）and urinary tract calculi（13 patients），with five patients exhibiting extra-renal manifestations such as renal tubular acidosis and hyperaldosteronism. Stone composition analysis identified calcium oxalate（16 patients），cystine（4 patients），and carbonate apatite（1 patient）. Metabolic abnormalities were prevalent，including hypocitraturia（11 patients），hyperoxaluria（8 patients），and hypercalciuria（7 patients），with eight patients presenting two or more concurrent disorders. All patients underwent WES and comprehensive metabolic evaluation. Sequencing was performed on an Illumina Hiseq4000 platform，achieving a mean depth of > 100× and coverage of > 98% in target regions. Variants were classified according to the American College of Medical Genetics and Genomics（ACMG）guidelines.Results:WES identified 12 distinct genes across autosomal recessive（9 genes： AGXT， GRHPR， ATP6V1B1， SLC12A1， KCNJ1， SLC3A1， SLC7A9， SLC34A3， WFS1），autosomal dominant（2 genes： CASR， ADCY10），and X-linked recessive（1 gene： CLCN5）inheritance patterns. Genotype-phenotype correlations revealed mutations associated with primary hyperoxaluria（8 patients），hypercalciuria（7 patients），and renal malformation due to a WFS1 mutation（1 patient）. A positive genetic diagnosis was achieved in 100% of patients with either urinary oxalate > 1 000 μmol/24 h or cystine stones. 8 patients received a diagnosis of hereditary stone disease at their first presentation（non-delayed group），while 13 experienced a mean diagnostic delay of（9.6 ± 3.9）years. The delayed diagnosis group had a significantly older age at initial stone onset［（10.2 ± 5.3）years vs.（6.8 ± 3.1）years， P = 0.03］and a higher incidence of impaired renal function（6 patients vs. 1 patient， P = 0.04）. Analysis of diagnostic delay by gene subgroup showed delays in 2/4 patients with cystinuria［ SLC3A1/ SLC7A9；（8.2 ± 3.5）years］，5/8 with primary hyperoxaluria［ AGXT/ GRHPR；（10.5 ± 4.1）years］，5/7 with hypercalciuria-related genes［ CASR/ ADCY10/ SLC12A1/ KCNJ1/ SLC34A3；（9.8 ± 3.8）years］，and 1/2 with other genes［ ATP6V1B1/ WFS1/ CLCN5；（7.6 ± 2.2）years］. Among 32 mutation sites detected，21 were classified as pathogenic/likely pathogenic and 11 as variants of uncertain significance. Four novel mutations were identified： ATP6V1B1（presenting with renal tubular acidosis，nephrocalcinosis，and hypocitraturia）， WFS1（presenting with renal malrotation，hydronephrosis，and stones without metabolic abnormalities）， SLC12A1（presenting with Bartter syndrome type 1，chronic renal insufficiency，hypercalciuria，hypocitraturia，alkalosis，and hyperaldosteronism），and SLC3A1（presenting with bilateral renal stones and cystinuria）. Conclusions:WES is crucial in identifying the underlying etiology of urolithiasis and can guide targeted treatment. We recommend early WES for patients with an initial stone presentation before age 25，those with nephrocalcinosis，or those with abnormal metabolic workups to facilitate precise diagnosis and preventive care.

Objective:To further study on the biological characteristics of vaccinia virus VG9 strain obtained by passaging from vaccinia virus Tiantan strain(VTT).Methods:The freeze-dried vaccinia virus VG9 strain was reconstituted and inoculated into Vero cells for recovery and sequential passaging. The strain was identified by indirect immunofluorescence and PCR amplification. The whole genome of VG9 was sequenced using next-generation sequencing technology. The sequencing results were compared with the reference sequence of VTT and the genome sequences of other 33 orthopoxviral strains, and a phylogenetic tree was drawn. A purified clone of VG9, namely VG9-V3-3, was obtained by terminal dilution method. Preliminary studies were conducted to characterize the biological properties of this clone, such as virus titer and intradermal virulence in rabbit.Results:VG9 could specifically bind to the rabbit antibody of VTT by indirect immunofluorescence identification, and the PCR amplification results proved that this strain contained the characteristic TK gene fragment of vaccinia virus. Whole-genome sequencing showed that the genome length of VG9 was 183 596 bp, including 165 150 bp in coding region. The sequencing results were compared with the NCBI core nucleotide database, and showed that the sequences of VG9 and multiple existing Tiantan isolates were highly homologous (consistency>99.5%). Sequence comparison revealed a total of 749 nucleotide site differences between the gene sequences of VG9 and TP5 isolate of VTT. The viral titer of the VG9-V3-3 isolate was 6.3 lg (PFU/ml), and the rabbit intradermal virulence assay showed that its virulence was significantly weakened compared with that of VTT.Conclusions:The biological characteristics of vaccinia virus VG9 strain are preliminarily studied and a purified clone strain named VG9-V3-3 is obtained. The viral titer of the isolate is basically stable, and the virulence is significantly weakened compared with that of VTT. In-depth study on the immunogenicity of VG9-V3-3 will be carried out in the future to explore its feasibility as a vaccine production strain.

Objective:To further study on the biological characteristics of vaccinia virus VG9 strain obtained by passaging from vaccinia virus Tiantan strain(VTT).Methods:The freeze-dried vaccinia virus VG9 strain was reconstituted and inoculated into Vero cells for recovery and sequential passaging. The strain was identified by indirect immunofluorescence and PCR amplification. The whole genome of VG9 was sequenced using next-generation sequencing technology. The sequencing results were compared with the reference sequence of VTT and the genome sequences of other 33 orthopoxviral strains, and a phylogenetic tree was drawn. A purified clone of VG9, namely VG9-V3-3, was obtained by terminal dilution method. Preliminary studies were conducted to characterize the biological properties of this clone, such as virus titer and intradermal virulence in rabbit.Results:VG9 could specifically bind to the rabbit antibody of VTT by indirect immunofluorescence identification, and the PCR amplification results proved that this strain contained the characteristic TK gene fragment of vaccinia virus. Whole-genome sequencing showed that the genome length of VG9 was 183 596 bp, including 165 150 bp in coding region. The sequencing results were compared with the NCBI core nucleotide database, and showed that the sequences of VG9 and multiple existing Tiantan isolates were highly homologous (consistency>99.5%). Sequence comparison revealed a total of 749 nucleotide site differences between the gene sequences of VG9 and TP5 isolate of VTT. The viral titer of the VG9-V3-3 isolate was 6.3 lg (PFU/ml), and the rabbit intradermal virulence assay showed that its virulence was significantly weakened compared with that of VTT.Conclusions:The biological characteristics of vaccinia virus VG9 strain are preliminarily studied and a purified clone strain named VG9-V3-3 is obtained. The viral titer of the isolate is basically stable, and the virulence is significantly weakened compared with that of VTT. In-depth study on the immunogenicity of VG9-V3-3 will be carried out in the future to explore its feasibility as a vaccine production strain.

OBJECTIVE: To carry out genetic testing for a XXY fetus suggested by non-invasive prenatal testing (NIPT). METHODS: G-banding karyotyping, fluorescence in situ hybridization (FISH) and chromosomal microarray analysis (CMA) were performed on amniocytes from the fetus. The genitalia of the fetus was also examined by Doppler ultrasonography. The result was verified with peripheral blood samples from its parents and a brother. RESULTS: The fetus was found to have a 46,XX karyotype. CMA showed presence of sequences from Yp11.2 (2.635 Mb) and Yp11.31p11.2 (3.706 Mb). FISH assay suggested that the SRY fragment on Yp has translocated to Xpter. No karyotypic or pathogenic CNVs was detected in its parents and brother. The fetus was ultimately diagnosed with 46,XX (SRY positive) male syndrome. CONCLUSION The combination of G-banding karyotyping, FISH, and CMA is of great significance for attaining accurate prenatal diagnosis for this fetus.

Objective: To assess the value of non-invasive prenatal testing (NIPT) for the identification of fetal chromosomal aneuploidies. Methods: For 9470 pregnant women with a moderate-to-high risk by conventional serological screening or advanced maternal age, peripheral venous blood samples were collected and, following extraction of free fetal DNA, subjected to large-scale parallel sequencing on a Illumina Hiseq2000 platform. Those with a high risk by NIPT were validated by invasive prenatal diagnosis. Results: Out of the 9470 samples, 194 cases (2.0%) were positive by NIPT testing. These included 50 trisomy 21, 11 trisomy 18, 17 trisomy 13, 44 other autosomal aneuploidies, 55 sex chromosomal aneuploidies, and 17 chromosomal copy number variations. As validated by amniotic fluid or umbilical blood chromosomal karyotyping analysis, NIPT has a false positive rate of 2.0%, 18.2%, 41.2%, 97.7%, 81.8%, 94.1%, respectively. The test has a sensitivity of 100% and a specificity of 98.79%. Conclusion For common chromosomal aneuploidies such as trisomy 21 and trisomy 18, NIPT has a good sensitivity and specificity, therefore has good value for clinical application.

OBJECTIVETo assess the value of combined chromosomal karyotyping and BACs-on-Beads(BoBs) assay for the prenatal diagnosis of high risk gravida from Ningbo.

METHODSFor 2779 women, results of conventional karyotyping analysis and BoBs assay were compared.

RESULTSFor common aneuploidies involving chromosomes 13, 18, 21, X and Y, the two methods have yielded a concordance rate of 98.78%. Eight cases detected with microduplication by BoBs were missed by karyotyping analysis. On the other hand, 17 structural chromosomal abnormalities, 10 chimeras and 1 triploidy detected by karyotyping analysis were missed by BoBs.

CONCLUSIONThe BoBs technology has featured high throughput and rapidity, and can detect 9 microdeletion syndromes, which can improve the quality of prenatal diagnosis and provide an ideal complementary for conventional chromosomal karyotyping.

Adult ; Chromosome Aberrations ; Chromosome Deletion ; Chromosomes, Artificial, Bacterial ; genetics ; Female ; Humans ; Karyotyping ; methods ; Pregnancy ; Prenatal Diagnosis ; methods

This study was aimed to evaluate the clinical effect and safety of Qing-Fei Tong-Luo (QFTL) ointment for treating children with pneumonia.Randomized controlled trial (RCT) was conducted among 460 cases of children with pneumonia.The observation group was given QFTL ointment combined with basic treatment.And the control group was only treated by basic treatment.Evaluation was given on the total clinical efficacy,disappeared time of fever,cough,expectoration,shortness of breath,and medication safety.The incidence of respiratory diseases was followed up on the 30th days after drug withdrawal.The results showed that in the aspect of clinical efficacy between two groups,the cure rate of the observation group was 98.26％,and that of the control group was 93.89％,with statistic significance (P ＜ 0.05).The cure rate of the observation group was better than that of the control group.There was statistical difference on expectoration disappeared time (P ＜ 0.05).There was no statistical difference on disappeared time of fever,cough and shortness of breath (P ＞ 0.05).There was statistical difference on the incidence of respiratory diseases on the 30th days followed-up after drug withdrawal (P ＜ 0.05).There was no statistical difference on the incidence of upper respiratory tract infection,pneumonia and asthma (P ＞ 0.05).No adverse reactions occurred in the observation group.It was concluded that QFTL ointment combined with basic therapy on the treatment of pneumonia in children was significantly better than the control group in the aspect of clinical efficacy,expectoration disappeared time and the incidence of bronchitis.It is safe and effective.The prognosis is good and worthy of promotion in the clinical practice.