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本文对1例60岁男性轻度认知障碍、头晕患者的临床症状、影像学和基因检测结果进行分析。该患者既往有高血压、高脂血症、高尿酸血症病史，以及长期吸烟、饮酒史，其母亲曾患痴呆，头颅MRI示多发腔隙性脑梗死、脑白质变性和多发微出血病灶，基因检测报告示HTRA1基因c.1174T>C杂合突变，以上结果符合HTRA1基因杂合突变相关遗传性脑小血管病的诊断。 Abstract: Clinical symptoms, imaging findings, and genetic testing results of a 60-year-old male patient with mild cognitive impairment and dizziness were analyzed. The patient had a history of hypertension, hyperlipidemia, hyperuricemia, and long-term smoking and alcohol consumption. His mother had dementia. Cranial magnetic resonance images revealed multiple lacunar infarctions, white matter degeneration, and multiple microbleeding lesions. Genetic testing report showed a heterozygous mutation in the HTRA1 gene (c.1174T > C). All of above results are consistent with the diagnosis of heterozygous HTRA1-related hereditary cerebral small vessel disease.

BACKGROUND: From the pathological mechanism of Western medicine, ligamentum flavum hypertrophy is the key pathogenic factor of lumbar spinal stenosis, and there is a lack of biological information on lumbar spinal stenosis of the Qi deficiency and blood stasis type. OBJECTIVE: To analyze and compare differential protein expression between hypertrophic and normal ligamentum flavum in patients with lumbar spinal stenosis of the Qi deficiency and blood stasis type. METHODS: Ligamentum flavum tissue samples were collected from six lumbar spinal stenosis patients with Qi deficiency and blood stasis, including three cases of ligamentum flavum hypertrophy (experimental group) and three cases of normal ligamentum flavum (control group). 4D Label free quantitative proteomic detection was performed to screen differentially expressed proteins. Gene oncology and Kyoto Encyclopedia of Genes and Genomes were used for enrichment analysis. RESULTS AND CONCLUSION: There were 183 differentially expressed proteins between the two groups, including 87 up-regulated and 96 down-regulated. Gene oncology enrichment analysis showed that biological processes mainly focused on cell processes, biological regulation and response to stimuli. Cell composition was concentrated in cell, intracellular, and protein-complex species. The main molecular functions included linkage, catalytic activity and molecular function regulator. The up-regulated proteins were mainly enriched to lysosomal signaling pathway, rheumatoid arthritis signaling pathway, and Staphylococcus aureus infection signaling pathway, while the down-regulated proteins were enriched to eight signaling pathways, namely p53 signaling pathway, renal cell carcinoma signaling pathway, transforming growth factor β signaling pathway, ubiquitin-mediated protein hydrolysis signaling pathway, Ca signaling pathway, cGMP-PKG signaling pathway, hypoxia-inducible factor 1 signaling pathway, and proteoglycan signaling pathway in cancer. INHBA, MMP14, TNC, HTRA1, FGF2 were the important differentially expressed proteins in the experimental group. To conclude, there are differential protein expressions between hypertrophic and normal ligamentum flavum in patients with Qi-stagnation and blood-stasis type lumbar spinal stenosis. INHBA may be the determinant of this disease, and its mechanism may be the activation of transforming growth factor β/Smad related signaling pathway, causing ligamentum flavum hypertrophy and subsequently leading to lumbar spinal stenosis. [ABSTRACT FROM AUTHOR]

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Objective This study aimed at describing the frequency of rare variants of monogenic cerebral small vessel diseases (CSVD) in a cohort of patients with CSVD, and to explore its clinical relevance. Methods This study included CSVD patients visiting the Neurology Department of Peking Union Medical College Hospital(PUMCH) from March 2017 to January 2022, collecting their demographic and clinical information and DNA samples for whole-exome sequencing. Descriptive analysis and statistical analysis were conducted exploring the differences between monogenic CSVD-related gene mutation carriers and noncarriers. Results A total of 292 patients were included, 51.03% of whom carried one or more rare variants of monogenic CSVD-related genes. The most common rare low-frequency variants were located in the NOTCH3 gene (70 patients, 23.97%), followed by HTRA1 and COL4A1/COL4A2 (22 patients, 7.53%) respectively. Among the subgroup of patients without a family history of stroke (n=176), the frequency of rare variants was as high as 47.16%. Compared with non-carriers, the carriers were diagnosed at a younger age (58.76±13.71 vs. 63.46±13.21, P=0.003). No difference was found in phenotypes among single-SNP carriers, multiple-SNPs carriers, and noncarriers. Conclusions The frequency of rare mutation of monogenic CSVD-related genes were relatively high in Chinese CSVD cohort. The most common rare variant was within the NOTCH3, followed by HTRA1 and COL4A1/COL4A2 genes. For CSVD patients of unknown causes, genetic screening should not be neglected even if there is not a family history of the disease.

Objective To explore the role of HTRA1 in the pathogenesis of keloids. Methods The expression of HTRA1 was detected by real time PCR, Western blot and immunohistochemistry. The dynamic changes of TGF-β1 activity were measured by luciferase reporting assay. Results Compared to normal skin, the expression of HTRA1 and TGF-β1 in keloids was significantly elevated. Co-culture with recombinant human HTRA1 (rhHTRA1) significantly increased the activation level of TGF-β1 and the phosphorylation level of Smad2. Conclusion HTRA1 was involved in the pathogenesis of keloids through regulating the activation of latent TGF-β1 in keloid fibroblasts. Therefore, HTRA1 can promote the development of keloids. [ABSTRACT FROM AUTHOR]

Objective: To construct the lentiviral vector containing HTRA1 as well as its mutant gene(HTRA1-Mut,1091TC) and express them in HBVSMCs. Methods: HTRA1 and HTRA1-Mut gene regional fragment were amplified by RT-PCR and then cloned into the plasmid GV287.Lentiviral plasmid, p Helperl.0, and p Helper2.0 were co-transfected into 293 T cells to obtain recombinant virus. The lentiviral titer was detected. Collected lentivirus was applied to infect human vascular smooth muscle cells. Results: The recombinant lentiviral vectors were constructed and were confirmed by PCR and sequencing analysis. They could efficiently transfect 293 T cells and express in the cells. The lentiviral titer was 2E+8 TU/ml. HBVSMCs produced fluorescent expression after lentivirus transfection, and HTRA1 gene expressed over 80%. Transfected HBVSMCs grew in order without inducing excessive death rate. Conclusion: HTRA1 and HTRA1-Mut recombined lentiviruses with high viral titer were successfully constructed and packaged, and the HTRA1 and HTRA1-Mut gene could be transfected into HBVSMCs with stable and high expression in infected cells, and these cells might be applied for mechanism researches of HTRA1 gene. [ABSTRACT FROM AUTHOR]

Objective To construct the plasmid pSilencer4.1/HtrA1 and stably transfect human trophoblastic cell line HPT-8. Methods We detected the expression of HtrA1 in cell line HPT-8 with immunohistochemical SABC, constructed the plasmid pSilencer4.1/HtrA1, transfected human trophoblastic cell line HPT-8 with plasmid pSilencer4.1/HtrA1 and negative plasmid pSilencer4.1, and observed the cell expression after transfection. Results About 90% of cellular cytoplasm of HPT-8 was dyed brown while the nucleus was negatively stained. Selective concentration of G418 was 200μg/mL for HPT-8 to be transfected pSilencer4.1/HtrA1. With RT-PCR and Western blot methods, the expression of HPT-8 transfected plasmid Psilence4.1/HtrA1 was significantly downregulated compared with that of cells with negative plasmid and normal cells (P<0.05). There was no significant difference in absorbance value between blank plasmid group and HPT-8 group (P>0.05). Conclusion We successfully constructed stable trophoblastic cell line HPT-8/ pSilencer4.1-HtrA1 with silenced expression of HtrA1, which lays foundation for further research. [ABSTRACT FROM AUTHOR]