Genotype-phenotype pattern analysis of pathogenic PAX9 variants in Chinese Han families with non-syndromic oligodontia.

Background: Non-syndromic oligodontia is characterized by the absence of six or more permanent teeth, excluding third molars, and can have aesthetic, masticatory, and psychological consequences. Previous studies have shown that PAX9 is associated with autosomal dominant forms of oligodontia but the precise molecular mechanisms are still unknown. Methods: Whole-exome and Sanger sequencing were performed on a cohort of approximately 28 probands with NSO, for mutation analysis. Bioinformatic analysis was performed on the potential variants. Immunofluorescence assay, western blotting, and qPCR were used to explore the preliminary functional impact of the variant PAX9 proteins. We reviewed PAX9-related NSO articles in PubMed to analyze the genotype-phenotype correlations. Results: We identified three novel PAX9 variants in Chinese Han families: c.152G>T (p.Gly51Val), c.239delC (p.Thr82Profs*3), and c.409C>T (q.Gln137Ter). In addition, two previously reported missense variants were identified: c.140G>C (p.Arg47Pro) and c.146C>T (p.Ser49Leu) (reference sequence NM_006194.4). Structural modeling revealed that all missense variants were located in the highly conserved paired domain. The other variants led to premature termination of the protein, causing structural impairment of the PAX9 protein. Immunofluorescence assay showed abnormal subcellular localizations of the missense variants (R47P, S49L, and G51V). In human dental pulp stem cells, western blotting and qPCR showed decreased expression of PAX9 variants (c.140G>C, p.R47P, and c.152G>T, p.G51V) compared with the wild-type group at both the transcription and translation levels. A review of published papers identified 64 PAX9 variants related to NSO and found that the most dominant feature was the high incidence of missing upper second molars, first molars, second premolars, and lower second molars. Conclusion: Three novel PAX9 variants were identified in Chinese Han families with NSO. These results extend the variant spectrum of PAX9 and provide a foundation for genetic diagnosis and counseling.

Novel PAX9 compound heterozygous variants in a Chinese family with non-syndromic oligodontia and genotype-phenotype analysis of PAX9 variants.

OBJECTIVE: Studies have reported that >91.9% of non-syndromic tooth agenesis cases are caused by seven pathogenic genes. To report novel heterozygous PAX9 variants in a Chinese family with non-syndromic oligodontia and summarize the reported genotype-phenotype relationship of PAX9 variants. METHODOLOGY: We recruited 28 patients with non-syndromic oligodontia who were admitted to the Hospital of Stomatology Hebei Medical University (China) from 2018 to 2021. Peripheral blood was collected from the probands and their core family members for whole-exome sequencing (WES) and variants were verified by Sanger sequencing. Bioinformatics tools were used to predict the pathogenicity of the variants. SWISS-MODEL homology modeling was used to analyze the three-dimensional structural changes of variant proteins. We also analyzed the genotype-phenotype relationships of PAX9 variants. RESULTS: We identified novel compound heterozygous PAX9 variants (reference sequence NM_001372076.1) in a Chinese family with non-syndromic oligodontia: a new missense variant c.1010C>A (p.T337K) in exon 4 and a new frameshift variant c.330_331insGT (p.D113Afs*9) in exon 2, which was identified as the pathogenic variant in this family. This discovery expands the known variant spectrum of PAX9; then, we summarized the phenotypes of non-syndromic oligodontia with PAX9 variants. CONCLUSION: We found that PAX9 variants commonly lead to loss of the second molars.

Observation and Analysis of Clinical Efficacy of Zhuang Medicine Lotus Acupuncture Cupping Stasis Therapy on Patients with Postherpetic Neuralgia.

OBJECTIVE: To explore the clinical efficacy of Zhuang Medicine Lotus Acupuncture Cupping Stasis Therapy on patients with postherpetic neuralgia (PHN) and its action mechanism. METHODS: 36 patients are randomly divided into Lotus Acupuncture Cupping Stasis Therapy group, pure cupping group and gabapentin group, with a total of five observation points for the first, fifth, tenth, fifteenth, and twentieth sessions of therapy (one session every three days). At each observation point, the venous blood of the patients is taken, and the contents of and changes in WNT3a, Frizzled8, ß-catenin, IL-18, TNF-α, NR2B, NK-1 and SP are tested by ELISA, RT-PCR and WesternBlot, respectively. The VAS scores and safety of the patients in the three groups are compared. RESULTS: With increased time spent in therapy, the VAS scores of patients in each group decreased gradually and there was a significant reduction in pain in patients in the Lotus Acupuncture Cupping Stasis Therapy group compared to the gabapentin and pure cupping groups (P<0.05). The levels of IL-18, TNF-α, NK-1, SP, WNT3a, Frizzled 8 and ß-catenin in the serum of all patients experienced a constant decline over time (P<0.05); the levels of the aforesaid factors in the serum of patients in the Lotus Acupuncture Cupping Stasis Therapy group dropped remarkably after the tenth session of therapy compared to those in gabapentin and pure cupping groups (P<0.05). CONCLUSIONS: Zhuang Medicine Lotus Acupuncture Cupping Stasis Therapy can significantly reduce the pain of PHN patients, with a good therapeutic effect, and it is worthy of clinical use.

Novel PAX9 compound heterozygous variants in a Chinese family with non-syndromic oligodontia and genotype-phenotype analysis of PAX9 variants

Abstract Studies have reported that >91.9% of non-syndromic tooth agenesis cases are caused by seven pathogenic genes. Objective To report novel heterozygous PAX9 variants in a Chinese family with non-syndromic oligodontia and summarize the reported genotype-phenotype relationship of PAX9 variants. Methodology We recruited 28 patients with non-syndromic oligodontia who were admitted to the Hospital of Stomatology Hebei Medical University (China) from 2018 to 2021. Peripheral blood was collected from the probands and their core family members for whole-exome sequencing (WES) and variants were verified by Sanger sequencing. Bioinformatics tools were used to predict the pathogenicity of the variants. SWISS-MODEL homology modeling was used to analyze the three-dimensional structural changes of variant proteins. We also analyzed the genotype-phenotype relationships of PAX9 variants. Results We identified novel compound heterozygous PAX9 variants (reference sequence NM_001372076.1) in a Chinese family with non-syndromic oligodontia: a new missense variant c.1010C>A (p.T337K) in exon 4 and a new frameshift variant c.330_331insGT (p.D113Afs*9) in exon 2, which was identified as the pathogenic variant in this family. This discovery expands the known variant spectrum of PAX9; then, we summarized the phenotypes of non-syndromic oligodontia with PAX9 variants. Conclusion We found that PAX9 variants commonly lead to loss of the second molars.

[Overview of "tian-yin-yang" acupuncture, an acupuncture school of Huang's Zhuang medicine of Guangxi].

The theory of "tian-yin-yang" of Zhuang medicine was explored and the academic achievements and experience of professor HUANG Jin-ming were summarized to explain the theory and clinical characteristics of tian-yin-yang acupuncture, an acupuncture school of Huang 's Zhuang medicine of Guangxi. This acupuncture method is guided by the theory of "three-qi synchronization" and "tian-yin-yang" of Zhuang medicine, based on the theory of "three channels and two paths", with regulating qi as the method and regulating spirit as the basis. After the patient is calmed and resting, the micro needle shallow needling technique is adopted, mainly at the umbilical ring point, so as to achieve the purpose of regulating the mind and treating the root cause.

[Efficacy on nervous tinnitus of kidney deficiency treated with Zhuang medicine at Qineihuan point and conventional acupuncture therapy].

OBJECTIVE: To compare the difference of the clinical efficacy in treatment of nervous tinnitus of kidney deficiency between the combined therapy of Zhuang medicine at Qineihuan point combined with the conventional acupuncture and simple conventional acupancture. METHODS: Sixty patients were randomized into an observation group and a control group, 30 cases in each one. In the control group, the conventional acupuncture was applied to Taixi (KI 3), Zhaohai (KI 6), Tinggong (SI 19) and Waiguan (TE 5), etc. and the needles were retained for 30 min. In the observation group, on the basic treatment as the control group, Zhuang medicine acupuncture at Qineihuan point was added. The treatment was given once every day, 10 treatments made one session and there were 2 days of interval between the sessions. In 3 sessions of treatment, the changes of tinnitus were observed and the clinical efficacy was evaluated. RESULTS: After treatment, tinnitus score and tinnitus grade were all improved as compared with those before treatment in the two groups (all P<0. 05) and the results in the observation group were better than those in the control group (all P<0. 05). The curative and remarkably effective rate was 63. 3% (19/30) and the total effective rate was 93. 3% (28/30) in the observation group, better than 30. 0% (9/30) and 73. 3% (22/30) in the control group (both P<0. 05). CONCLUSION: The combined therapy of Zhuang medicine at Qineihuan point and conventional acupuncture achieves the better efficacy on nervous tinnitus of kidney deficiency as compared with the simple conventional acupuncture.

[Umbilical ring acupoints and its clinical application in Zhuang medicine].

The theoretical basis, location, belonging of zang-fu, treatment function and indications, applying principle and manipulation of Umbilical Ring acupoints in Zhuang medicine are explained in this paper. According to Zhuang medicine, umbilicus is an epitome of the body and all the zang-fu and organs in the body have corresponding epitomes like a fetus in front-standing position. The umbilicus is not only a micro-diagnosis system, but also a window for illness treatment that could be divided into superficial, middle and deep layer to respectively communicate different zang-fu and organs. The umbilical inner ring and outer ring are collectively called Umbilical Ring acupoints, they could dredge paths, regulate the balance of qi and blood to regulate qi, expel poison, tonify deficiency and remove stasis to treat many types of diseases in the whole body.