Objective To screen small molecule inhibitors of Fusobacterium nucleatum （Fn） based on commercially available compound libraries, and investigate their anti-colorectal cancer activities under Fn intervention in order to obtain novel anti-colorectal cancer lead compounds. Methods The promotion of colorectal cancer proliferation on organoid was validated by Fn. Secondly, the effects of anti-Fn compounds on their in vitro anticancer activity under Fn’s co-incubation with colorectal cancer HCT116 cell were comparative investigated. Finally, in vivo anticancer efficacy of highly active compounds on nude mouse colon cancer HCT116 transplanted tumor under the intervention of Fn was evaluated by gavage. Results Fn could significantly promote the proliferation of rectal cancer organoids. 9 anti-Fn active compounds could significantly enhance their in vitro anticancer activity under Fn’s co-incubation with HCT116 cells. Methotrexate had the strongest anti-cancer activity with IC₅₀ as 0.03 μmol/L. The combined use of methotrexate (0.5 mg/kg) and PD-1 (5.0 mg/kg) had a stronger anti-tumor effect than their standalone use. Conclusion As new small molecule inhibitor of Fn, methotrexate exhibited good in vitro and in vivo anti-colorectal cancer activity against HCT116 cells and nude mouse xenografts under Fn intervention, which showed the foundation for subsequent structural optimization, and could be expected to expand the new indications of methotrexate.

ObjectiveThis study aimed to evaluate the clinical efficacy of Ruyi Zhenbaowan（RYZBW）in the treatment of initial and early knee osteoarthritis （KOA） through a prospective multicenter，randomized，double-blind，and placebo-parallel controlled trial. MethodFrom October 13^th， 2021 to December 25^th， 2021， 240 KOA subjects meeting the acceptance criteria were enrolled in 15 sub-centers including Wangjing Hospital， Chinese Academy of Chinese Medical Sciences， and they were randomly divided into observation group and control group， with 120 cases in each group. The intervention measures for the observation group were RYZBW + health education， and the intervention measures for the control group were RYZBW placebo + health education. The intervention period in both groups was four weeks， and they were followed up for four weeks after the intervention. The primary outcome measure was the total score of Western Ontario and McMaster University Osteoarthritis Index score （WOMAC score）， and the secondary outcome measures were the response rate of visual scale （VAS） pain score， WOMAC sub item scores （joint pain， joint stiffness， and joint function）， quality of life （SF-12） score， and traditional Chinese medicine （TCM） syndrome score. Result（1） Efficacy evaluation. The marginal model results showed that the observation group was better than the control group in improving the WOMAC total score and WOMAC pain score in the treatment of KOA with RYZBW， and the difference was statistically significant （P<0.05）. There was no significant difference between the two groups in improving VAS score response rate， WOMAC function score， WOMAC stiffness score， SF12-PCS （quality of life-physical health） score， SF12-MCS （quality of life-mental health） score， and TCM syndrome score. （2） Subgroup analysis. ① In terms of VAS score response rate， the response rate of the observation group was higher than that of the control group for subjects with baseline VAS score of （4， 5］， and the difference was statistically significant （P<0.05）. ② In terms of TCM syndrome score， for subjects aged ［56， 60］ and ［61， 65］， the decrease in total TCM syndrome score in the observation group was better than that in the control group， and the difference was statistically significant （P<0.05）. ConclusionTibetan medicine RYZBW has good clinical efficacy in improving WOMAC total score， VAS score response rate， WOMAC pain score， WOMAC function score， and TCM syndrome score for patients with initial and early KOA， which can fill the lack of Tibetan medicine RYZBW in the treatment of KOA and make a demonstration study for the inheritance and development of ethnic medicine.

Objective:To investigate the risk factors of non-valvular paroxysmal atrial fibrillation (NVPAF) with cerebral ischemic stroke(CIS) and analyze NVPAF by using left atrial automatic imaging (AFILA). Logistic regression model was established for left atrial(LA) function parameters.Methods:A total of 205 patients with NVPAF were included in the study and divided into the NVPAF group without ischemic stroke (154 patients) and the CIS group (51 patients). The clinical baseline data, blood biochemical results and AFILA ultrasound data of all patients were collected. Univariate analysis was performed to compare the above data between the two groups of patients. The independent risk factors were obtained by multivariate logistic regression analysis. Logistic regression model was compared with CHA2DS2-VASc scoring system in terms of area under ROC curve, sensitivity and specificity.Results:There were significant differences in age, CHA2DS2-VASc score, taking anticoagulant drugs, history of hypertension, diabetes and coronary heart disease, LAEF, S_R, S_CT, WBC, NEUT, HCY, UREA, NDD, NT-proBNP, Fibrinogen(Fib), Cardiac troponin I(cTnI) and NLR between the two groups (all P<0.05). The results of multifactor analysis showed that: age, hypertension, S_ CT, UREA, NLR, Fib and cTnI were independent risk factors associated with CIS in patients with paroxysmal atrial fibrillation[ OR value: 1.608 ( P=0.003), 3.821 ( P=0.019), 1.259 ( P=0.001), 1.326( P=0.001), 1.352 ( P=0.011), 1.502 ( P=0.042), 7.651( P=0.001)]. After adjusting for the age, sex and history of hypertension included in CHA2DS2-VASc score, S_CT significantly led to NVPAF complicated with stroke[ OR value 1.259 (1.095-1.447), P=0.001]. The diagnostic efficacy of Logistic regression model is better than that of CHA2DS2-VASc scoring (AUC of 0.931 vs 0.717, 95% CI: 0.896-0.967 vs 0.634-0.799, sensitivity of 0.883 vs 0.755, specificity of 0.849 vs 0.713, all P<0.001). Conclusions:Age, hypertension, S_CT, UREA, NLR, fibrinogen, cTnI are independently associated risk factors for patients with combined CIS; The diagnostic efficacy of Logistic regression model is better than that of CHA2DS2-VASc scoring model.And the sensitivity and specificity are high.

KRAS‒PDEδ interaction is revealed as a promising target for suppressing the function of mutant KRAS. The bottleneck in clinical development of PDEδ inhibitors is the poor antitumor activity of known chemotypes. Here, we identified novel spiro-cyclic PDEδ inhibitors with potent antitumor activity both in vitro and in vivo. In particular, compound 36l (K _D = 127 ± 16 nmol/L) effectively bound to PDEδ and interfered with KRAS-PDEδ interaction. It influenced the distribution of KRAS in Mia PaCa-2 cells, downregulated the phosphorylation of t-ERK and t-AKT and promoted apoptosis of the cells. The novel inhibitor 36l exhibited significant in vivo antitumor potency in pancreatic cancer patient-derived xenograft (PDX) models. It represents a promising lead compound for investigating the druggability of KRAS‒PDEδ interaction.

Objective: To investigate the pathogenicity of rare damaging variants from ciliary pathway to human lumbosacral neural tube defects(NTDs). Methods: The coding region sequences of 49 ciliary genes were amplified by AmpliSeq technique and sequenced by PGM sequencing platform for screening the rare damaging variants, and the parents without phenotype were used as controls to evaluate the pathogenicity of variants. Results: The rare de novo mutation of GLI3 gene(c.C580 T,p.H194 Y) was detected in one patient.The rare complex heterozygous mutations of CRB2 gene(c.G1392 C,p.R464 S;c.T3448 C,p.C1150 R) was detected in another patient. Conclusion Rare damaging variants from ciliary pathway may be associated with the occurrence of human lumbosacral NTDs.

Objective:To compare the efficacy of bilayer artificial dermis graft plus single layer dermal template and bilayer artificial dermis graft only in repairing lower extremity wounds with large area of exposed bone.Methods:A retrospective case-control study was conducted to analyze the clinical data of 34 patients with 37 wounds of the lower extremity involving large area of exposed bone admitted to Beijing Jishuitan Hospital from November 2009 to November 2020. There were 27 males and 7 females, aged 9-67 years [35.5(29, 45)years]. The exposed bone in the lower leg, ankle and foot was greater than 10 cm 2 in size (the shortest distance from edge to edge of bony exposure was more than 2 cm). At the first stage, the wounds were grafted with bilayer type artificial dermis only for 21 wounds of 20 patients in Group A, and grafted with bilayer type artificial dermis plus single layer dermal template for 16 wounds of 14 patients in Group B. At the second stage, the auto-skin graft was performed in the two groups. The wound healing rate was observed in all patients, and was compared between the two groups at 2 weeks and 2 months after the second stage operation. At the same time, the interval between first stage and second stage surgery was measured. The Vancouver Scar Scale (VSS) was used to evaluate the scar in the skin grafting area in the two groups at 5-6 months after the second stage operation. Results:All patients were followed up for 1 to 24 months [5(2, 7.5)months]. The total excellent and good wound healing rate in all patients was 81%(30/37) at 2 weeks and 97%(36/37) at 2 months. There was no significant difference between the Group A and Group B in the excellent and good wound healing rate at 2 weeks [(76%(16/21) vs. 88%(14/16)] and at 2 months [95%(20/21) vs. 100%(16/16)] ( P>0.05). In Group A, the bilayer artificial dermis was grafted into 4 wounds again to complete exposed bone coverage. However, all wounds in Group B were covered initially without re-grafting. The interval between the two-stage operation was 20(16, 21)days in Group A after the 4 patients who underwent secondary artificial dermal transplantation were excluded, showing no significant difference from 21(21, 23)days in Group B ( P>0.05). At 5-6 months after the second stage operation, the VSS score in Group B [(8.0±1.2)points] was significant less than that in Group A [(9.2±1.1)days] ( P<0.05). In the sub-index of VSS, the score of color and softness of scar in Group B [(2.0±0.6)points, (1.6±0.5)points] were significantly improved compared to those in Group A [(2.5±0.5)points, (2.2±0.7)points] ( P<0.05). Conclusions:The artificial dermis grafting is effective in treatment of lower extremity wounds with large area of exposed bone. However, the bilayer artificial dermis graft plus single layer dermal template can avoid artificial dermal re-graft in repair of large area of exposed bone, and the interval between two-stage operation is not significantly prolonged. Moerover, the color and texture of scar after skin grafting and wound repair efficiency and quality are improved.

A great challenge in multi-targeting drug discovery is to identify drug-like lead compounds with therapeutic advantages over single target inhibitors and drug combinations. Inspired by our previous efforts in designing antitumor evodiamine derivatives, herein selective histone deacetylase 1 (HDAC1) and topoisomerase 2 (TOP2) dual inhibitors were successfully identified, which showed potent and antitumor potency. Particularly, compound was orally active and possessed excellent antitumor activity in the HCT116 xenograft model (TGI = 75.2%, 150 mg/kg, .) without significant toxicity, which was more potent than HDAC inhibitor vorinostat, TOP inhibitor evodiamine and their combination. Taken together, this study highlights the therapeutic advantages of evodiamine-based HDAC1/TOP2 dual inhibitors and provides valuable leads for the development of novel multi-targeting antitumor agents.

OBJECTIVE: To detect pathogenic variation in a pedigree affected with hereditary spastic paraplegia type 31 and explore its molecular pathogenesis. METHODS: Customized Roche NimbleGen capture probes were used to capture all exons of the target genes in relation with hereditary spastic paraplegia. The DNA samples were also assayed with fluorescent quantitative PCR as well as chromosomal microarray analysis using CytoScan HD chip. RESULTS: The proband and her father and grandfather were found to carry a deletion for position 85 992 693-86 842 693 on chromosome 2, which spanned approximately 900 kb and encompassed the REEP1 gene. The latter has been specifically associated with hereditary spastic paraplegia type 31. The same deletion was not found in her mother who is phenotypically normal. CONCLUSION The deletional variation of the REEP1 gene probably underlies the disease in this pedigree.
Female ; Humans ; Membrane Transport Proteins ; supply & distribution ; Paraplegia ; Pedigree ; Sequence Deletion ; Spastic Paraplegia, Hereditary ; genetics

Natural product evodiamine and its derivatives represent a promising class of multi-target antitumor agents. However, the clinical development of these compounds has been hampered by a poor understanding of their antitumor mechanisms. To tackle this obstacle, herein, novel fluorescent probes were designed to elucidate the antitumor mode of action of 10-hydroxyevodiamine. This compound was proven to be distributed in the mitochondria and lysosomes and to act by autophagy and apoptosis mechanisms.

OBJECTIVE To explore the origin and morphological features of small supernumerary marker chromosomes (sSMCs) in Turner syndrome. METHODS For 5 cases of Turner syndrome with a sSMC identified by conventional G-banding, dual-color fluorescence in situ hybridization (FISH) was applied to explore their origin and morphological features. RESULTS Among the 5 cases, 3 have derived from the X chromosome, which included 2 ring chromosomes and 1 centric minute. For the 2 sSMCs derived from the Y chromosome, 1 was ring or isodicentric chromosome, while the other was an isodicentric chromosome. CONCLUSION The sSMCs found in Turner syndrome have almost all derived from sex chromosomes. The majority of sSMCs derived from the X chromosome will form ring chromosomes, while a minority will form centric minute. While most sSMC derived from Y chromosome may exist as isodicentric chromosomes, and a small number may exist as rings. For Turner syndrome patients with sSMCs, dual-color FISH may be used to delineate their origins to facilitate genetic counseling and selection of clinical regime.