OBJECTIVETo detect potential mutations of COL1A1 and COL1A2 genes with polymerase chain reaction-high-resolution melting analysis(PCR-HRMA) in a proband diagnosed with osteogenesis imperfecta (OI).

METHODSPeripheral blood samples were collected from the proband and members of his family as well as healthy controls. The mutations were detected by PCR-HRMA and confirmed by direct sequencing. Potential effects of the mutations were predicted using softwares including PolyPhen, SIFT and Align GVGD.

RESULTSThe PCR-HRMA has indicated mutations in exon 45 of the COL1A1 gene in the proband as well as his parents, which were presented as the difference in the melting curves between the patients and the control samples. Sequencing analysis confirmed that the proband has carried two heterozygous mutations (c.3235G>A, p.Gly1079Ser and c.3247G>A, p.Ala1083Thr) in exon 45 of the COL1A1 gene. Among them, c.3235G>A was predicted to have impeded alpha helix structure domain, which was inherited from the father who also had OI. c.3247G>A was inherited from mother who had a normal phenotype. All three softwares predicted that the c.3235G>A mutation can interfere with the function of the protein, while the c.3247G>A may have a benign effect by PolyPhen analysis.

CONCLUSIONThe study identified two mutations (c.3235G>A and c.3247G>A) occurred simultaneously in COL1A1 gene in a case. The case is the first reported in human collagen mutation database. As identified,mutation of c.3235G>A may be the major cause of the disease in the proband.

Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Case-Control Studies ; Child ; Child, Preschool ; China ; Collagen Type I ; genetics ; Exons ; Female ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Osteogenesis Imperfecta ; genetics ; Pedigree ; Point Mutation

Objective: To explore the expression of Dickkopf-3(DKK-3) mRNA in oral squamous cell carcinoma(OSCC) and the relationship between the promoter methylation status and the carcinogenesis of OSCC. Methods: The expression of DKK-3 gene in 51 cases of OSCC and corresponding normal mucosa tissue was detected by PT-PCR and methylation-specific PCR, respectively. The relationship between DKK-3 and the clinical pathological features of the patients was analyzed with SPSS 13. 0. Results: The expression level of DKK-3 in OSCC group was lower than that in the control(t =-12. 580, P＜ 0. 05). The methylation rate of DKK-3 gene promoter region in OSCC group was significantly higher than that in the control(χ2 = 19. 273, P＜ 0. 05). The mRNA expression level of DKK-3 gene in OSCC with methylation group was lower than that in the control(t =-2. 817, P＜ 0. 05). Conclusion: Down-regulation of DKK-3 gene expression and hypermethylation of promoter region are important mechanisms in the pathogenesis of OSCC. The hypermethylation of DKK-3 promoter may be the main cause of transcriptional silencing.

OBJECTIVETo explore genetic mutations and clinical features of osteogenesis imperfecta type V.

METHODSClinical record of five patients (including one familial case) with osteogenesis imperfecta type V were retrospectively analyzed. Peripheral blood samples of the patients, one family member, as well as healthy controls were collected. Mutation of IFITM5 gene was identified by PCR amplification and Sanger sequencing.

RESULTSA heterozygous mutation (c.-14C>T) in the 5-UTR of the IFITM5 gene was identified in all of the patients and one mother. The clinical findings included frequent fractures and spine and/or extremities deformities, absence of dentinogenesis imperfecta, absence of hearing impairment, and blue sclera in 1 case. Radiographic findings revealed calcification of the interosseous membrane between the radius-ulna in all cases. Hyperplastic callus formation was found in 3 cases. Four had radial-head dislocation.

CONCLUSIONA single heterozygous mutation c.-14C>T was found in the 5-UTR of the IFITM5 gene in 5 patients with osteogensis imperfecta type V. The patients showed specific radiological features including calcification of interosseous membrane, hyperplastic callus formation, and radial-head dislocation.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Humans ; Male ; Mutation ; Osteogenesis Imperfecta ; diagnostic imaging ; genetics ; Young Adult

Objective To summarize the clinicopathological characteristics and prognostic factors of salivary duct carcinoma(SDC)patients.Methods This study was reviewed and approved by the Ethics Committee,and informed consent was obtained from the patients.The clinical data of 30 SDC patients who were admitted to the Fourth Hospital of Hebei Medical University from 2014 to 2022,including case records,pathological diagnoses,immunohistochemical indicators,treatment methods,follow-up data,and other data,were retrospectively analyzed.SPSS 26.0 software was used to process the data and construct relevant curves.The chi-square test was used to analyze the correlation between different immunohistochemical indices and the recurrence and metastasis of SDC,and a single factor was used to ana-lyze clinical prognostic factors.Results Among the 30 SDC patients,the male-to-female ratio was 5∶1,with a median age of 61.5 years.Approximately 60％of cases occurred in the parotid gland,whereas the remainder occurred in the submaxillary gland,sublingual gland,or minor salivary gland.Among them,19 patients were androgen receptor-positive,23 patients were human epidermal growth factor receptor-2 positive,and 26 patients were Ki-67 positive.Postoperative follow-up was 18-94 months,with a median follow-up of 37 months.There were 13 cases of recurrence and 14 cases of distant metastasis.The 5-year overall survival rate was only 31.2％.The long-term survival of patients who underwent postoperative radiotherapy and chemoradiotherapy was better than that of patients who underwent surgery alone(P=0.027).T stage and lymph node invasion were associated with prognosis and survival(P＜0.05).There was a correlation between a Ki-67-positive cell count ≥ 40％and postoperative recurrence or metastasis(P=0.025).Conclusion Radi-cal surgery combined with postoperative radiotherapy and chemoradiotherapy is helpful for improving long-term overall survival,and tumor T stage and lymph node metastasis may be the main factors affecting the prognosis of patients with SDC.Patients with Ki-67-positive cell counts ≥ 40％are prone to postoperative recurrence or metastasis.

Oral static biofilm model is an important tool for in vitro simulation of oral microecological environment, which has become an important method for studying the pathogenesis of various oral diseases and testing the efficacy of various drugs, oral care products and foods due to its low cost, high throughput, good reliability and easy operation. The establishment of oral static biofilm models allows the selection of different devices, inoculum sources, media, substrates and culture conditions according to the purpose of the study, and the evaluation of biofilm growth by various methods such as measuring biomass, metabolic activity, community structure and performing visualization analysis. This paper summarizes the methodological elements reported in recent years for the establishment and evaluation of oral static biofilm models, and analyzes and discusses the applicability of various methods in the hope of contributing to the research and production practice in related fields.
Biofilms ; Reproducibility of Results

ObjectiveTo investigate the possible mechanism of Hewei Anshen Formula （和胃安神方） in the treatment of insomnia. MethodsSixty male SD rats were randomly divided into the normal group， the model group， the eszopiclone group and the low-， medium- and high-dose Hewei Anshen Formula groups. The insomnia model was constructed by intraperitoneal injection of p-chlorophenylalanine （PCPA） for 2 days in all groups except the normal group. After successful modelling， the eszopiclone group was given 0.33 mg/（kg·d） eszopiclone aqueous solution by gavage， the low-， medium- and high-dose Hewei Anshen Formula groups were given 10 ml/kg of Hewei Anshen Formula with a concentration of 1， 2 and 4 g/ml， respectively， and the rats in the normal group and the model group were given 10 ml/kg of saline by gavage， once a day for 7 consecutive days. The general condition of the rats was observed during the experiment， and the body mass of the rats was measured every day after medication administration. The following day after the last medication administration， pentobarbital sodium co-test was used to observe the sleep condition， and the sleep latency and sleep duration were recorded； immunohistochemistry and Western blot were used to detect the expression of hypothalamic clock rhythm regulating protein （CLOCK） and brain and muscle aromatic hydrocarbon receptor nuclear transporter-like protein 1 （BMAL1） in the rats. ResultsThe body mass of rats in the model group was lower than that of rats in the normal group at all time points （P＜0.01）； compared with the same time in the eszopiclone group， the body mass of rats in the low-dose Hewei Anshen Formula group was elevated on the 5th， 6th and 7th days of medication administration （P＜0.05）. Compared with the normal group， the sleep duration of rats in the model group was shortened （P＜0.01）； compared with the model group， the sleep duration of rats in each dosage group increased （P＜0.01）， and the difference between the high-dose Hewei Anshen Formula group and the eszopiclone group showed no statistically significant （P＞0.05）， while the sleep duration of the low- and medium-dose Hewei Anshen Formula groups were shorterned than the eszopiclone group （P＜0.01）. The difference in sleep latency showed no statistically significant among each group （P＞0.05）. The results of both immunohistochemistry and Western blotting showed that the expression of CLOCK and BMAL1 in the hypothalamus of rats in the model group was significantly reduced compared with that in the normal group （P＜0.01）； the expression of CLOCK and BMAL1 in the hypothalamus of rats in the low- and high-dose Hewei Anshen Formula groups increased than that in the model group （P＜0.05 or P＜0.01）. ConclusionHewei Anshen Formula can improve insomnia in model rats， and its mechanism of action may be related to the up-regulation of the expression of the hypothalamic biological clock genes CLOCK and BMAL1 protein.