Objective·To analyze the root canal diameter and taper of mandibular first molars using cone-beam CT(CBCT),investigate age-related changes in these parameters,and propose optimized clinical protocols for root canal preparation.Methods·From October 2022 to October 2023,CBCT images of 240 healthy mandibular first molars(120 three-canal type cases and 120 four-canal type cases)were collected from patients aged 20?59 years at the Stomatology Center,Haikou Affiliated Hospital of Central South University Xiangya School of Medicine.The three-canal type and the four-canal type samples were respectively divided into four age groups(20?29,30?39,40?49,and 50?59 years),with 30 cases in each group.Root canal length was measured,and buccolingual and mesiodistal diameters were recorded at four levels(coronal,middle,apical,and foramen).The tapers of the coronal 1/3,middle 1/3,and apical 1/3 segments were calculated in both buccolingual and mesiodistal directions.Differences among the age groups were compared.Results·The buccolingual diameters of all root canals exceeded the corresponding mesiodistal diameters.At several levels of most root canals,the diameters in the<50-year-old groups were significantly larger than those in the 50?59-year-old group(P<0.05).The buccolingual tapers exceeded the corresponding mesiodistal tapers in all root canals of mandibular first molars.The mean mesiodistal tapers in different age groups ranged from 0.07 to 0.34 in the coronal 1/3 segment,0.03 to 0.09 in the middle 1/3 segment,and 0.05 to 0.11 in the apical 1/3 segment.At several levels of most root canals,the tapers in the<50-year-old groups were significantly larger than those in the 50?59-year-old group(P<0.05).Conclusion·Mandibular first molars exhibit age-related narrowing of root canal diameter and reduced taper in individuals aged≥50 years.For initial apical file selection,#15 files are recommended for the distal canals in the three-canal type,while#10 files are advised for all other canals in the three-canal type and all canals in the four-canal type.For patients aged 50 years or older,the files for the distal canals in the three-canal type can be adjusted to#10,and the files for the mesiobuccal and distallingual canals in the four-canal type to#8.Regarding preparation taper,a 0.08-taper orifice opener is recommended for the coronal 1/3 segment.During mid-lower segment preparation,0.04-taper master apical files are suggested for the mesiobuccal and mesiolingual canals in the three-canal type and the mesiolingual and distallingual canals in the four-canal type,0.06-taper files for the distal canals in the three-canal type and the mesiobuccal canals in the four-canal type,and 0.08-taper files for the distobuccal canals in the four-canal type.Root canal obturation is recommended to be performed using vertical compaction with large-taper gutta-percha cones combined with bioceramic sealer.

Objective·To analyze the root canal diameter and taper of mandibular first molars using cone-beam CT(CBCT),investigate age-related changes in these parameters,and propose optimized clinical protocols for root canal preparation.Methods·From October 2022 to October 2023,CBCT images of 240 healthy mandibular first molars(120 three-canal type cases and 120 four-canal type cases)were collected from patients aged 20?59 years at the Stomatology Center,Haikou Affiliated Hospital of Central South University Xiangya School of Medicine.The three-canal type and the four-canal type samples were respectively divided into four age groups(20?29,30?39,40?49,and 50?59 years),with 30 cases in each group.Root canal length was measured,and buccolingual and mesiodistal diameters were recorded at four levels(coronal,middle,apical,and foramen).The tapers of the coronal 1/3,middle 1/3,and apical 1/3 segments were calculated in both buccolingual and mesiodistal directions.Differences among the age groups were compared.Results·The buccolingual diameters of all root canals exceeded the corresponding mesiodistal diameters.At several levels of most root canals,the diameters in the<50-year-old groups were significantly larger than those in the 50?59-year-old group(P<0.05).The buccolingual tapers exceeded the corresponding mesiodistal tapers in all root canals of mandibular first molars.The mean mesiodistal tapers in different age groups ranged from 0.07 to 0.34 in the coronal 1/3 segment,0.03 to 0.09 in the middle 1/3 segment,and 0.05 to 0.11 in the apical 1/3 segment.At several levels of most root canals,the tapers in the<50-year-old groups were significantly larger than those in the 50?59-year-old group(P<0.05).Conclusion·Mandibular first molars exhibit age-related narrowing of root canal diameter and reduced taper in individuals aged≥50 years.For initial apical file selection,#15 files are recommended for the distal canals in the three-canal type,while#10 files are advised for all other canals in the three-canal type and all canals in the four-canal type.For patients aged 50 years or older,the files for the distal canals in the three-canal type can be adjusted to#10,and the files for the mesiobuccal and distallingual canals in the four-canal type to#8.Regarding preparation taper,a 0.08-taper orifice opener is recommended for the coronal 1/3 segment.During mid-lower segment preparation,0.04-taper master apical files are suggested for the mesiobuccal and mesiolingual canals in the three-canal type and the mesiolingual and distallingual canals in the four-canal type,0.06-taper files for the distal canals in the three-canal type and the mesiobuccal canals in the four-canal type,and 0.08-taper files for the distobuccal canals in the four-canal type.Root canal obturation is recommended to be performed using vertical compaction with large-taper gutta-percha cones combined with bioceramic sealer.

Objective:To simulate the neuroendoscopy assisted contralateral cervical 7 (C 7) nerve transfer via prespinal route and measure its relevant anatomical landmarks to explore the clinical feasibility and efficacy of this surgical approach for central upper limb spastic paralysis. Methods:(1) Six fresh cadaver specimens of the head and neck were obtained. Linear incisions of approximately 5 cm were made above the midpoint of the bilateral clavicles to simulate neuroendoscopy assisted contralateral C 7 nerve transfer via prespinal route. With the superior margin of midpoint of the clavicle as central point, distance to the distal bifurcation of the C 7 nerve, distances to the superior or inferior trunks of the proximal brachial plexus nerves, and distances to the exit of the intervertebral foramina of the C 6, C 7, and C 8 nerves were measured. (2) One patient with upper limb spastic paralysis after intracerebral hemorrhage accepted neuroendoscopy assisted contralateral C 7 nerve transfer via prespinal route; the clinical data and efficacy of the patient were retrospectively analyzed. Results:(1) The C 7 nerve, the upper trunk of brachial plexus formed by the C 5 and C 6 nerves and the lower trunk of brachial plexus formed by the C 8 and T 1 nerves could be exposed after neuroendoscopy assisted contralateral C 7 nerve transfer via prespinal route. The distance between the superior margin of midpoint of the clavicle and the distal bifurcation of the C 7 nerve is (2.20±0.11) cm, and its distance to the superior trunk of the proximal brachial plexus is (2.62±0.10) cm, and its distance to the inforior trunk of the proximal brachial plexus nerve is (2.72±0.11) cm. The distance between the superior margin of midpoint of the clavicle and the proximal C 7 nerve (at the exit of the intervertebral foramen) is (7.22±0.15) cm, its distance to the proximal C 6 nerve (at the exit of the intervertebral foramen) is (7.84±0.12) cm, and its distance to the proximal C 8 nerve (at the exit of the intervertebral foramen) is (6.96±0.12) cm. (2) The patient with central upper limb spastic paralysis accepted neuroendoscopy assisted contralateral C 7 nerve transfer via prespinal route successfully, with surgical time lasting for 2 h and bleeding amount of 20 mL. After surgery, the incision healed well, and the patient experienced pain and numbness in the healthy side of the upper limb with subsided symptoms one month after surgery. The spasticity symptoms of the affected upper limb obviously improved after surgery with decreased muscle tone. Follow-up after discharge was performed for 14 months, and the muscle strength of the affected upper limb recovered to level 1 +. Conclusion:Neuroendoscopy assisted contralateral C 7 nerve transfer via prespinal route can expose the proximal and distal C 7 nerves, with minimal invasion; this clinical study has preliminarily confirmed the safety and effectiveness of this transfer via prespinal route in central upper limb spastic paralysis.

Objective:To investigate whether interleukin(IL)-1β is involved in pyroptosis which leads to mouse islet β cell line βTC-6 cell damage, and to explore the role of JNK inhibitor SP600125 in inhibiting IL-1β induced βTC-6 cell pyroptosis.Methods:βTC-6 cell line and mouse islets were incubated with IL-1β for 48 h or intervened with both JNK inhibitor SP600125 and IL-1R antagonist IL-1Ra, then GSDMD expression and β cell pyroptosis morphology were detected by immunofluorescence staining of GSDMD and DAPI. The expression levels of Gsdmd, IL-1β and IL-18 mRNAs were detected by real time fluorescence PCR, and apoptosis was examined by Annexin-V/7-AAD staining combined with flow cytometry.Results:βTC-6 cell pyroptotic body was significantly increased in the IL-1β treated group compared with the control group, and the expressions of pyroptosis related genes Gsdmd, IL-1β, and IL-18 mRNA were significantly higher( P<0.05), and apoptosis was increased, suggesting that IL-1β effectively induced the βTC-6 cell pyroptosis, IL-1Ra prevented IL-1β induced βTC-6 cell pyroptosis. In the presence of JNK inhibitor SP600125, IL-1β treatment failed to induce the expressions of Gsdmd and IL-18 mRNA, markers of pyroptosis, and reduced the rate of apoptosis, indicating that SP600125 suppressed IL-1β induced βTC-6 cell pyroptosis. Conclusion:Pyroptosis is one of the mechanisms of βTC-6 cell impairment caused by IL-1β, and SP600125, a JNK inhibitor, can block the IL-1β induced pyroptosis pathway and has a potential role in inhibiting βTC-6 cell pyroptosis.

Objective To explore the mutation characteristics of neurexin 1(NRXN1)gene in children with Tourette syndrome(TS).Methods A total of 524 children with TS were enrolled.DNA extracted from peripheral blood was sequenced for NRXN1 gene by using target region sequencing which was further verified by using Sanger sequencing.DNAMAN software,SIFT,PolyPhen2,Mutation Taster,FATHMM and ClinPred were used to analyze the hazard of suspected variants.Finally,the genotype and phenotype of the patients with NRXN1 gene variants were analyzed.Results We found 13 variants of the NRXN1 gene in 13 TS patients such as 11 point mutations and 2 deletion mutations including two novel point mutations:c.79G>T(p.A27S)and c.58G>T(p.G20C).The other nine point mutations and two deletion mutations were c.3523A>G(p.I1175V),c.4180A>T(p.T1394S),c.1697A>T(p.H566L),c.3715G>A(p.A1239T),c.878A>C(p.N293T),c.475C>T(p.P159S),c.320C>T(p.T107M),c.365A>G(p.Q122R),c.611T>A(p.L204Q)c.68_79del(p.G23_G26del),c.65_79del(p.G22_G26del).Bioinformatics analysis showed that the six gene variants c.58G>T,c.1697A>T,c.475C>T,c.365A>G,c.878A>C,c.79G>T were relatively harmful.There were 6 children with different parts of the tic,1 child with obsessive-compulsive symptoms,1 child with emotional instability,3 children with irritability,6 children did not have repetitive language,attention deficit,hyperactivity disorder,sleep disorder and depression.Conclusion NRXN1 gene mutation sites are detected in TS children,which expands the NRXN1 mutation spectrum.Children with different gene variants exhibit different clinical manifestations and the relationship between genotype and phenotype need further exploration.

Objective:To investigate the association between gene polymorphisms in vitamin D receptor(VDR) and Tourette syndrome (TS).Methods:The genetic contributions of VDR FokI (rs2228570), BsmI (rs1544410), and Cdx2 (rs11568820) polymorphisms were genotyped by TaqMan allelic discrimination real-time (RT)-PCR, which evaluated by a case-control analysis in 417 TS patients and 442 healthy controls, and followed by a family-based study in 417 TS trios.Chi-square test and relative risk analysis were conducted by IBM SPSS 23.0 software.Results:FokI (rs2228570) had three genotypes(CC=109, CT=235, TT=73); BsmI (rs1544410) had three genotypes(AA=2, AG=45, GG=370); Cdx2 (rs11568820) had three genotypes(AA=71, AG=200, GG=146). No significant difference in genotype ( χ2=5.516, P=0.063; χ2=3.466, P=0.177; χ2=0.561, P=0.755, respectively) or allele frequencies( χ2=0.840, P=0.359; χ2=3.376, P=0.066; χ2=0.051, P=0.822, respectively)of FokI, BsmI and Cdx2 were identified between TS patients and control groups.No significant over-transmission was identified for these three polymorphisms among 417 TS trios in the family-based study (TDT for FokI: χ2=0.009, P=0.962; for BsmI: χ2=1.220, P=0.320; and for Cdx2: χ2=0.260, P=0.646). Haplotype relative risk (HRR) analysis and haplotype-based haplotype relative risk (HHRR) analysis showed no significant difference in allele frequencies distribution of FokI, BsmI and Cdx2 (all P>0.05). Conclusion:VDR receptor gene polymorphism has no effect on TS susceptibility in the Chinese Han population. However, a potential role of VDR should be explored in more polymorphisms, different populations and larger samples.

Although non-invasive prenatal testing has been widely used, it has certain limitations. As the gold standard of prenatal diagnosis, G-banding karyotype analysis is time-consuming and laborious. Fluorescence in situ hybridization (FISH), as a method for detecting samples with non-radioactive signals, does not require cell culture and has a short turnover time, and can diagnose aneuploidies of chromosomes 13, 18, 21, X, Y with efficiency, which can solve the problems such as insufficient testing ability and long diagnosis period for karyotype analysis. To standardize the procedures of prenatal FISH assay and enhance laboratory quality management, the Expert Committee of the Prenatal Screening and Diagnosis Laboratory of the Clinical Test Center of the National Health Commission and the Inter-laboratory Quality Assessment Committee of the Neonatal Genetic and Metabolic Disease Screening Laboratory have formulated this consensus.

Objective To investigate the features and characteristics of GLIS3 gene mutation in patients with congenital hypothyroidism(CH),and to establish the theoretical basis for gene diagnosis and prenatal diagnosis of CH.Methods Genomic DNA was extracted from peripheral blood leukocytes of 50 patients with CH who were collected from February 2007 to November 2016 in Shandong Province.The exon 2 to 11 of GLIS3 were amplified with 11 pairs of sequence specific primers designed by Primer 5.0.Polymerase chain reaction and the first generation of sequencing method(Sanger sequencing) were used to detect the mutation.Comparison of the sequencing results with the GLIS3 reference sequence (National Center for Biotechnology Information Reference Sequence:NC_000009.12) helped to screen gene mutations.Results The 50 CH patients included 22 boys and 28 girls,and the sex ratio was 1.0 ∶ 1.3.The mean age was (2.5 ± 0.5) years.Six cases (12％) had thyroid gland hypoplasia,23 cases (46％) had thyroid gland agenesis and 21 cases(42％) with ectopic thyroid gland.C2507A missense mutation was found in exon 10 of GLIS3 in a thyroid gland agenesis case,which might result in proline to glutamine substitution at codon 836.One mutant (rs780019691,c.C289T) was detected which was nonsense mutation (Arg→Stop) in another thyroid gland agenesis child.Conclusions The mutation rate of GLIS3 gene is very low in CH children of Shandong province.Further studies are needed to investigate the relationship between GLIS3 genotypes and clinical phenotypes.

Objective To explore the association between the GRIN3B gene and Tourette syndrome (TS) in children by screening mutations in the coding region of this gene.Methods Fifty-one children with TS and their parents in the Affiliated Hospital of Qingdao University from October 2015 to November 2016 were selected as an experimental group,41 cases of boys,and 10 cases of girls,aged 6-16 years[(9.78 ±3.64)years],while 60 people aged 22-45 years in the health examination center were selected in the control group,49 cases were male,1 1 cases were female,aged 22-45 years [(29.08 ± 2.89) years].DNA was extracted from 51 patients with TS,their parents and 60 controls.PCR was applied to amplify the encoding region of GRIN3B gene and Sanger sequencing was used to sequence,then GRIN3B sequencing results were compared with the NCBI gene encoding region sequence (NM_138690.2)to test whether these patients carried gene mutation and to verify the findings from their family.Results c.C460T gene variant of GRIN3B was found in 2 patients (p.P154S);c.T1187C (p.L396S) variant of GRIN3B gene was found in 10 patients and both of abnormal GRIN3B sites lead to changes in amino acid.The 2 peak sequencing maps were obtained by Sanger sequencing but nothing was found in their parents.Conclusion The mutation of GRIN3B gene may be related to the development of TS.

Muhidrug resistance (MDR) is one of the important reasons for the failure of clinical anticancer drugs,involving multiple mechanisms.Among them,the classical MDR mechanism mediated by P-glycoprotein (P-gp) is closely related to the formation of MDR,which can excrete intracellular chemotherapeutic drugs through the "drug pump" effect and significantly reduce the therapeutic effect.Curcumin is mainly extracted from the underground rhizome of Chinese medicine turmeric,with a wide range of pharmacological activity.Recent studies have found that curcumin also has a role in reversing the MDR of the tumor,by inhibiting both P-gp function and expression,and this process involves a variety of signal paths.