Clinical and genetic characteristics of 9 rare cases with coexistence of dual genetic diagnoses / 中华儿科杂志

Objective: To analyze the clinical and genetic characteristics of pediatric patients with dual genetic diagnoses (DGD). Methods: Clinical and genetic data of pediatric patients with DGD from January 2021 to February 2022 in Peking University First Hospital were collected and analyzed retrospectively. Results: Among the 9 children, 6 were boys and 3 were girls. The age of last visit or follow-up was 5.0 (2.7,6.8) years. The main clinical manifestations included motor retardation, mental retardation, multiple malformations, and skeletal deformity. Cases 1-4 were all all boys, showed myopathic gait, poor running and jumping, and significantly increased level of serum creatine kinase. Disease-causing variations in Duchenne muscular dystrophy (DMD) gene were confirmed by genetic testing. The 4 children were diagnosed with DMD or Becker muscular dystrophy combined with a second genetic disease, including hypertrophic osteoarthropathy, spinal muscular atrophy, fragile X syndrome, and cerebral cavernous malformations type 3, respectively. Cases 5-9 were clinically and genetically diagnosed as COL9A1 gene-related multiple epiphyseal dysplasia type 6 combined with NF1 gene-related neurofibromatosis type 1, COL6A3 gene-related Bethlem myopathy with WNT1 gene-related osteogenesis imperfecta type XV, Turner syndrome (45, X0/46, XX chimera) with TH gene-related Segawa syndrome, Chromosome 22q11.2 microduplication syndrome with DYNC1H1 gene-related autosomal dominant lower extremity-predominant spinal muscular atrophy-1, and ANKRD11 gene-related KBG syndrome combined with IRF2BPL gene-related neurodevelopmental disorder with regression, abnormal movement, language loss and epilepsy. DMD was the most common, and there were 6 autosomal dominant diseases caused by de novo heterozygous pathogenic variations. Conclusions: Pediatric patients with coexistence of double genetic diagnoses show complex phenotypes. When the clinical manifestations and progression are not fully consistent with the diagnosed rare genetic disease, a second rare genetic disease should be considered, and autosomal dominant diseases caused by de novo heterozygous pathogenic variation should be paid attention to. Trio-based whole-exome sequencing combining a variety of molecular genetic tests would be helpful for precise diagnosis.

Association of genetic variants of hTERT with susceptibility to hepatocellular carcinoma / 中国现代普通外科进展

Objective:To explore the correlation between several single-nucleotide polymorphisms (SNPs) loci on the gene of human telomerase reverse transcriptase (hTERT)(including rs2736100、rs2853676) and the susceptibility to Hepatocellular carcinoma in Qingdao area.Methods:This case-control study included 165 diagnosed Hepatocellular carcinoma,195 with chronic hepatitis B and 402 Healthy people.Polymerase chain restriction fragment length polymorphism were performed to analyze the genotype of the loci on the gene of hTERT.Results:The frequency of TT genotype on rs2736100 were higher in the Hepatocellular carcinoma group compared to the control group、chronic hepatitis B and no HCC people(P＜0.05).The additive odds ratio of TT genotype for the risk of Hepatocellular carcinoma was 2.13、2.17 and 2.14.The frequency of AA genotype on rs2853676 were higher in the Hepatocellular carcinoma group compared to the control group、chronic hepatitis B and no HCC people (P＜0.05).The additive odds ratio ofTT genotype for the risk of Hepatocellular carcinoma was 3.42、2.39 and 3.02.Conclusion:There was a relationship between hTERT rs2736100 and rs2853676 polymorphism and susceptibility to Hepatocellular carcinoma.

Preliminary study on substitution of Antelope Horn in Danqi Piantan capsule with artificial bezoar / 中国中药杂志

To study the protective effect of Danqi Piantan capsule ( DPC) and its antelope horn substitution (DPCAS) on the cerebral ischemia, in order to preliminary study the possibility of replacing antelope horn with artificial bezoar. In this study, the left middle cerebral artery occlusion (MCAO) was adopted. Totally 150 SD rats were randomly divided into 5 groups: the sham operation group, the model group, the Danqi Piantan capsule (DPC) group (0.246 g x kg(-1) x d(-1)), the Danqi Piantan capsule without antelope horn (DPCRA) group (0.246 g x kg(-1) x d(-1)), the Danqi Piantan capsule without antelope horn and with double artificial bezoar (DPCDB) group (0.246 g x kg(-1) x d(-1)). The MCAO model was prepared 1 h later after the administration on the 5th day. At 24 h after the operation, the inner canthus blood was collected to determine the serum superoxide dismutase (SOD) activity and the endothelin (ET) content. At 72 h after the operation, the cerebral infarct size and the cerebral index were determined by TTC-staining. The fluorescent quantitative PCR method was used to detect brain Bcl-2, Caspase-3, IL-1β, P-selectin, E-selectin, ICAM-1 mRNA expressions. The mmunohistochemical method was used to detect ICAM-1, IL-1β, TNF-α, IL-6 expressions in ischemic penumbra. According to the results, compared with the model group, DPCDB and DPC groups showed almost consistent results, indicating both of the two group can significantly improved cerebral infarction index and cerebral index (P < 0.05), increase the serum SOD activity (P < 0.05), decrease the serum ET level and Caspase-3 expression, IL-1β, P-selectin, E-selectin, ICAM-1 mRNA expressions in brain tissues (P < 0.05) and expressions of ICAM-1, IL-1,6, TNF-α, IL-6 positive cells in ischemic penumbra (P < 0.05) and increase the Bcl-2 expression (P < 0.05). The DPCRA group showed much lower impacts on indexes than DPCDB and DPC groups. This suggests that DPCDB and DPC reveal similar efficacies and antelope horn in Danqi Piantan capsule can be substitutes by artificial bezoar.

Substitution of antelope horn in Danqi Piantan capsule with artificial bezoar in vitro / 中国中药杂志

The in vitro cell culture experiment was conducted to study the effect of Danqi Piantan capsule (DPC) and DPC dislodge the antelope horn with artificial bezoar double (DPCBD) on nerve regeneration and blood vessel regeneration and preliminarily investigate the possibility of substituting antelope horn in DPC with artificial bezoar. In this experiment, rats were randomly divided into 5 groups: the blank serum control group, the model group, DPC groups (0.306 g x kg(-1) x d(-1), the same below), DPC remove of antelope horn (DPCRA) groups and DPCBD groups. Brain microvascular endothelial cells cultured in vitro (BMEC), astrocytes and neural stem cells (NSC) were co-cultured to simulate neurovascular unit, label neurons with microtubule associated protein III (β-tubulin III) antibody and lable astrocytes with glial fibrillary acidicprotein (GFAP). ELISA was used for the detection of the content of BMEC lactate dehydrogenase instrument method (LDH), the inverted phase contrastmicroscope was adopted to observe the formation of BMEC tube like structure, the number of leukocytes and leukocytes adherent to BMEC were counted under the microscope, the expression levels of β-tubulin III and the ratio of GFAP positive cells was detected with inimmunofluorescence, and RT-PCR method was used to detect NGF, BDNF, VEGF and VEGFr-2 mRNA. According to the result, compared with the model group, both DPC and DPCBD can reduce LDH leakage, promote the formation of BMEC tube like structure, inhibit leukocytes and their adhesion to BMEC, increase the β-tubulin III positive cell differentiation proportion (P < 0. 01), reduce the proportion of GFAP positive cells (P < 0.01), increase the expressions of co-cultured NGF, VEGF, BDNF and VEGFr-2 mRNA to a certain extent, with the most significant difference on NGF and VEGF mRNA expressions (P < 0.05) and the same efficacy in both groups. DPCRA groups showed less impact on all indexes than that of DPCBD and DPC groups. The same efficacy of DPCBD and DPC on nerve regeneration and angiogenesis suggested that antelope horn in DPC can be substituted by artificial bezoar.

Effect of B7-H3 protein on the expression of macrophage-inflammatory protein 2 mRNA in Streptococcus pneumococcal meningitis mouse models / 中华实用儿科临床杂志

Objective To investigate the effect of B7-H3 protein,a collaborative signal molecule,on macrophage-inflammatory protein 2 (MIP-2) mRNA level in Streptococcus pneumococcal meningitis mouse models.Methods Forty-eight healthy male BALB/C mice at the age of 1.5-2.0 months were randomly divided into 4 groups:9 g/L saline group(NS group),B7-H3 protein group(B7-H3 group),Streptococcus pneumoniae group(SP group),and Streptococcus pneumoniae plus B7-H3 protein group(combination group),12 mice in each group.Mouse models were established by intracerebral ventricular injection with 9 g/L saline,B7-H3 protein,Streptococcus pneumoniae type 3 or Streptococcus pneumoniae type 3 plus B7-H3 protein.Neurobehavior of different groups was evaluated according to loeffler rule after injection for 6 h and 24 h,then the mice were sacrificed and MIP-2 mRNA levels were tested by Real-time PCR.The results were analyzed by SPSS 18.0 software.Results Neurobehavior scoring results showed that there were no significant differences between B7-H3 group and NS group (P ＞ 0.05) after infection for 6 h and 24 h,while the score of SP group was decreased compared with that of NS group (P ＜ 0.05),and the score of combination group was significantly decreased compared with that of SP group (P ＜ 0.05).Real-time PCR results showed that,compared with the NS group,the relative MIP-2 mRNA level in SP group increased after injection for 6 hours (1.210 ±0.932 vs 1.000 ± 0.008),but the difference was not significant (P ＞ 0.05),while at 24 h post infection,the relative MIP-2mRNA expressions in SP group were significantly increased compared with that of NS group(12.880 ± 7.792 vs 1.000 ±0.091),the difference was significant (P ＜ 0.05).At 6 h post infection,SP + B7-H3 treatment enhanced the MIP-2mRNA production compared to SP infection alone,but the difference was not significant [(1.240 ± 0.804) vs (1.210 ± 0.932)] (P ＞ 0.05) ; while at 24 h post infection,the difference was significant (38.760 ± 6.601 vs 12.880 ± 7.792) (P ＜ 0.05).Conclusion Collaborative signal molecule B7-H3 protein may increase MIP-2 mRNA level in Streptococcus pneumococcal meningitis mouse model,and exaggerate the clinical disease condition.