Objective To observe the value of self-made improved cone-beam CT(CBCT)fixation components for maxillofacial scanning.Methods CBCT fixation components(chin rest,temporal clamp)equipped by Planmeca ProMax 3D Mid dental panoramic X-ray system were modified to obtain improved CBCT fixation components,including improved chin rest,temporal clamp and occipital retention band.Totally 571 patients who underwent maxillofacial CBCT examinations for dental-related diseases were retrospectively enrolled,including 288 cases(group A)underwent scanning by a junior technician using original chin rest+original temporal clamp(component 1,n=78,A1 subgroup),modified chin rest+original temporal clamp(component 2,n=74,A2 subgroup),original chin rest+original temporal clamp+occipital retention band(component 3,n=68,A3 subgroup)or modified chin rest+original temporal clamp+occipital retention band(component 4,n=68,A4 subgroup),while the rest 283 cases(group B)underwent scanning by a senior using the above components,respectively,and were taken as B1(n=72),B2(n=69),B3(n=73)and B4(n=69)subgroups.The incidence of motion artifacts were compared among A2/B2,A3/B3,A4/B4 subgroups and A1/B1 subgroups.Results Compared with A1+B1 subgroups,the overall motion artifacts rate in A4+B4 subgroups decreased from 17.33％(26/150)to 8.03％(11/137)(P=0.019),while in A2+B2 subgroups(13.29％[19/143])and A3+B3 subgroups(9.93％[14/141])did not significantly decrease(P=0.337,0.067).In group A,no statistical difference of motion artifacts rate was found between A 2(17.57％[13/74])or A3(13.24％[9/68])subgroups and A1 subgroup(23.08％[18/78])(P=0.399,0.127),while the motion artifacts rate in A4 subgroup(10.29％[7/68])was lower than that in A1 subgroup(23.08％[18/78])(P=0.041).In group B,no significant difference of motion artifacts rates was found between B2,B3 or B4 subgroups(8.70％[6/69],6.85％[5/73],5.80％[4/69])and B1 subgroup(11.11％[8/72])(P=0.632,0.369,0.258).Conclusion Self-made improved CBCT fixation components could effectively secure patients' head and reduce motion artifacts in maxillofacial CBCT.

Objective To investigate the clinical features of spinocerebellar ataxia 12(SCA12).Methods The clinical data of a family with SC A1 2 diagnosed by gene detection were retrospectively analyzed,and the literature was reviewed.Results Three patients in this family were diagnosed as SCA12.Among them,2 patients with clinical symptoms were＞45 years old,the clinical symptoms were head tremor,upper limb tremor,voice tremor,dystonia and ataxia.Another one case was 31 years old,did not have relevant clinical manifestations yet.Cranial MRI showed that cerebral cortex atrophy was more serious than cerebellar atrophy in 2 patients.Conclusion SCA12 typically presents with upper limb tremor,with head tremor being less common,as the disease progresses,symptoms such as ataxia,dystonia,Parkinsonism,and cognitive impairment may develop,due to its highly heterogeneous clinical features,genetic testing can be helpful for diagnosis.

Objective To understand the current situation as well as the implementation of infection prevention and control measures against bacterial contamination of external and vaginal ultrasound probes in medical institutions(MIs),and explore the risk of ultrasound probe-related infection under current prevention and control measures.Methods Through on-site sampling,external ultrasound probes from 179 MIs and vaginal ultrasound probes from 83 MIs in 15 prefecture-level cities in Henan Province underwent quantitative bacterial detection,and the implemen-tation of relevant infection prevention and control measures was assessed using questionnaire survey.Results A to-tal of 513 ultrasound probe specimens were detected.Bacterial detection rate and exceedance rate of the disinfected vaginal ultrasound probes were 18.02％(20/111)and 2.70％(3/111),respectively,which were both lower than external ultrasound probes(40.05％[161/402]and 5.22％[21/402],respectively).Bacterial detection rate of dis-infected specimens was lower than that of the towel-wiping group([33.86％,149/440]vs[50.00％,27/54],x2=5.46,P=0.019).Bacterial detection rate of the quaternary ammonium salt disinfectant group was lower than that of the alcohol disinfectant group([17.31％,18/104]vs[40.63％,128/315],x2=18.74,P＜0.001).The quali-fied rates of specimens using different disinfection methods and with different probe types showed no statistically significant differences(all P＞0.05).The implementation rates of"single disinfection for every use"for external ul-trasound probes and vaginal ultrasound probes were 60.34％(108/179)and 70.54％(79/112),respectively.The disinfectants used were mainly quaternary ammonium salts(57.27％)and alcohols(36.36％).The coupling agents used for external and vaginal ultrasound examinations were mainly large packaged products for multiple time use,accounting for 93.85％and 57.89％,respectively.33.33％(56/168)of external ultrasound coupling agents and 20.00％(11/55)of vaginal ultrasound coupling agents were used for more than 7 days.In vaginal ultrasound exa-minations,the usage rates of disinfectant coupling agents and sterile coupling agents were 28.42％(27/95)and 14.74％(14/95),respectively.In vaginal ultrasound examinations,50.00％(56/112)of MIs used condoms as probe co-vers,and 45.54％(51/112)used sound-transmission ultrasound isolation membranes.60.55％(66/109)of MIs didn't change gloves after cleaning and disinfecting the vaginal ultrasound probe and before examination.Conclusion Ultrasound probes have cross infection risks in key processes such as reprocessing(disinfection/sterilization),phy-sical barrier use,coupling agent selection,and glove replacement.It is urgent to develop and implement standar-dized ultrasound probe infection prevention and control technical specifications based on evidence-based research.

Objective To examine the effects of SMAD4 on the proliferation and apoptosis of ovarian granulosa cells in rats with poly-cystic ovary syndrome(PCOS).Methods A PCOS rat model was established using DHEA,and ovarian granulosa cells were extracted and cultured in vitro.The expression of SMAD4 in ovarian granulosa cells was detected by quantitative real-time PCR and Western blot-ting.SMAD4-siRNA was transfected into ovarian granulosa cells from PCOS rats.The expression of SMAD4 mRNA after transfection was determined by quantitative real-time PCR.Western blotting was performed to detect the expression levels of PCNA,BAX,and BCL-2 proteins after transfection.A CCK-8 assay was performed to evaluate cell growth after siRNA interference.Results The HE staining results revealed that the number of ovarian follicular vacuoles increased and that the number of granulosa cell layers and corpus luteum decreased,thus indicating the establishment of a PCOS model.The FSHR positivity rate exceeded 95％.SMAD4 expression in ovarian granulosa cells was higher in the PCOS group than in the control group(P＜0.05).Furthermore,siRNA effectively reduced SMAD4 expression in ovarian granulosa cells of PCOS rats(P＜0.01),promoted proliferation,and inhibited the apoptosis of granulosa cells.Con-clusion The hindered growth of ovarian granulosa cells in PCOS rats may be linked to the overexpression of SMAD4 mRNA,which sug-gests that targeting SMAD4 could be a promising approach for treating ovulatory abnormalities in patients with PCOS.

Objective: To construct microRNA(miR)-22 gene knockout(miR-22-/-) mice using CRISPR/Cas 9 technology, to breed miR-22-/- mice and to identify their genotypes. Methods : In this experiment, CRISPR/Cas 9 technology was used to construct miR-22-/- genetically engineered mice. After gene identification, the F0 generation miR-22-/- mice were mated with wild-type mice in the same litter to obtain F1 generation miR-22-/- mice. The miR-22 knockout efficiency was analyzed at the RNA level by real-time fluorescence quantitative polymerase chain reaction(qPCR). Western blot was used to detect the interaction between miR-22 and target genes. Results : miR-22-/- mice were successfully constructed using CRISPR/Cas 9 technology, gene identification was performed on the bred mice, and three stable genotypes of miR-22+/+,miR-22+/-, and miR-22-/- were identified. The real-time fluorescence quantitative PCR detection results confirmed that miR-22-/- mice showed almost no expression of miR-22 in the heart, liver, lung, kidney, spleen, and thymus tissues compared to wild-type mice in the same litter. Western blot analysis showed that the relative expression level of NLRP3 protein in miR-22-/- mouse tissues was lower than that in wild-type mice. Conclusion A miR-22-/- mouse model is successfully constructed, and stable genetic homozygous miR-22-/- mice is obtained. This indicates that miR-22 has an inhibitory effect on the downstream target gene NLRP3.

Objective: To explore the breeding and genotyping of CCR2 knockout mice, and to verify the applicability of the polymerase chain reaction(PCR) method for genotype detection of CCR2 knockout mice. Methods: The introduced CCR2 pure male mice and wild-type female mice were mated and bred to produce the offspring generation, the obtained F1 generation heterozygous mice were continued to be mated. DNA was extracted by clipping the tail tissues of the mice at the age of 2 weeks, the target gene fragment was amplified by PCR, and the genotypic results were determined by agarose gel electrophoresis. The proportion of purebred progeny carrying the CCR2 knockout gene was increased by genetic crosses, the effect of CCR2 knockout in the progeny mice was verified by using Western blot against major immune cells and key organs, and flow cytometry was used to detect whether the knockout of the CCR2 gene had any effect on the function of the immune system by targeting the major immune cells. Results: CCR2 knockout mice were successfully bred and characterized, and three genotypes of F2 generation mice were obtained: CCR2+/+, CCR2+/-, and CCR2-/-. The offspring genotypes were identified by PCR, and Western blot showed extremely low CCR2 protein expression in CCR2 knockout mice. Flow analysis showed that CCR2 knockdown reduced the expression of CD4+T and Th1 cells in mouse spleen-derived T cells, but did not affect macrophage function. Conclusion Correct breeding and identification are important ways to get the pure CCR2 knockout mice, and PCR method for identifying mouse genotypes is simple, fast and reliable.

Objective To explore the diagnostic value of non-contrast chest CT in acute pulmonary thromboembolism(APTE).Methods A total of 187 patients with suspected APTE who underwent non-contrast chest CT and computed tomography pulmonary angiography(CTPA)within 2 h were included.Among 187 patients,there were 89 patients with APTE(APTE group)and 98 patients without APTE(control group).The clinical characteristics and chest CT features between the APTE group and the control group were compared.The sensitivity and specificity of non-contrast chest CT findings in the diagnosis of central APTE and peripheral APTE were analyzed.Results Chest CT showed pulmonary artery hyperdensity sign in 35 cases in the APTE group and none in the control group,with the difference was statistically significant(χ2=47.414,P<0.001);Subpleural shadow appeared in 33 cases in the APTE group and 11 cases in the control group,with the difference was statistically significant(χ2=17.327,P<0.001).The sensitivity and specificity of pulmonary artery hyperdensity sign in the diagnosis of central APTE and peripheral APTE were 72.92%,100%and 0%,100%,respectively.The sensitivity and specificity of sub-pleural shadow in the diagnosis of central APTE and peripheral APTE were 39.58%,88.78%and 34.15%,88.78%,respectively.The difference was not statistically significant in pleural effusion,pulmonary artery diameter,or pulmonary artery diameter to aorta diameter ratio between the two groups.Conclusion The pulmonary artery hyperdensity sign on non-contrast chest CT is a useful sign of APTE,which can avoid CTPA examination.

Objective:To elucidate the prognostic differences and risk factors associated with different patterns of recurrence and metastasis following pancreatic cancer (PC) resection.Methods:This is a retrospective case series study. Clinicopathological data and follow-up information were retrospectively collected from 210 patients who underwent surgery for PC at the Department of Hepatobiliary and Pancreatic Surgery, Peking University First Hospital, between January 2014 and December 2023. There were 114 males and 96 females; the age was (64.5±10.3) years (range: 29 to 89 years). Survival functions based on different times to recurrence and metastasis and different patterns of recurrence and metastasis were estimated using the Kaplan-Meier method, and survival differences among groups were compared using the Log-rank test. Identifying the optimal cutoff for time to postoperative recurrence/metastasis predicting overall survival (OS) in pancreatic cancer patients via the minimum p-value approach. Univariate and multivariate Cox regression analyses were performed to identify independent risk factors affecting OS following pancreatectomy.Results:A total of 210 patients met the inclusion and exclusion criteria. Among them, 71 patients remained free of recurrence and metastasis, while 139 developed recurrence and metastasis. The patterns included local recurrence ( n=34), liver metastasis ( n=39), lung metastasis ( n=11), peritoneal metastasis ( n=13), multiple sites metastasis ( n=38), bone metastasis ( n=3), and kidney metastasis ( n=1). OS was significantly shorter in the group with postoperative recurrence and metastasis compared to the group without recurrence/metastasis (23.07 months vs.not reached, P<0.01). The optimal cut-off time distinguishing early from late recurrence and metastasis was 13 months. There was a significant difference in post-recurrence survival between patients with early and late recurrence and metastasis (16.03 months vs. 52.40 months, P=0.009). The Kaplan-Meier survival curve showed that different postoperative recurrence and metastasis patterns had different impacts on OS, with lung metastasis showing the best prognosis compared to local recurrence, liver metastasis, peritoneal metastasis, and multiple sites metastasis ( P<0.01). Multivariate Cox analysis revealed that Eastern Cooperative Oncology Group (ECOG) score 1, postoperative carcinoembryonic antigen (CEA) ≥15 μg/L, poor tumor differentiation, postoperative local recurrence, liver metastasis, peritoneal metastasis, and multiple sites metastases are independent risk factors for postoperative recurrence and metastasis (all P<0.05). Conclusions:Considerable prognostic heterogeneity exists in postoperative PC patients depending on the site and pattern of recurrence or metastasis. Specifically, lung metastasis portends a significantly more favorable prognosis than liver metastasis, peritoneal metastasis, local recurrence, or multiple sites metastases. ECOG score 1, postoperative CEA≥15 μg/L, poor tumor differentiation, postoperative local recurrence, liver metastasis, peritoneal metastasis, and multiple sites metastases are independent risk factors for OS in postoperative PC patients.

Objective:To analyze the epidemiological characteristics and influencing factors of single-cigarette use and dual cigarette-cigar use in China.Methods:This study was a cross-sectional study that selected 85 638 urban and rural residents who met the inclusion criteria from the 2018 China Health Literacy Survey as research subjects. An analysis was conducted on 21 849 users of cigarettes and cigars among them. Due to the small number of individuals who exclusively used cigars (247 cases), the research subjects were divided into two categories: exclusive cigarette users and dual users of cigarettes and cigars. The groups were categorized by age (18-34 years, 35-54 years, ≥55 years), gender (male, female), education level (primary school and below, junior high school and high school, university and above) and annual household income (<20 000 yuan, 20 000-<80 000 yuan, ≥80 000 yuan) to compare the tobacco usage rate and conduct subgroup analyses for each subgroup. Multivariate logistic regression analysis was employed, incorporating general demographic characteristic information to explore the influencing factors of exclusive cigarette use and dual use of cigarettes and cigars, respectively.Results:The rate of exclusive cigarette use in our country was 24.3%, while the dual use rate of cigarettes and cigars was 0.9%. The exclusive cigarette use rate and the dual use rate of cigarettes and cigars among males were significantly higher than those among females (48.25% vs 2.48%, and 1.84% vs 0.06%) (both P<0.001). For males, the high-risk factors for exclusive cigarette use included living in urban areas ( OR: 1.37, 95% CI: 1.23-1.54), being Han ethnicity ( OR: 1.73, 95% CI: 1.51-1.98), and having an annual household income ≥20 000 yuan ( OR: 1.54, 95% CI: 1.38-1.82) while having a junior high school education or higher was a protective factor ( OR: 0.68, 95% CI: 0.52-0.90). Age≥35 years ( OR: 3.36, 95% CI: 2.62-4.32) and having a junior high school education or higher ( OR: 1.30, 95% CI: 1.02-1.67) were risk factors for dual use of cigarettes and cigars in males. Among females, living in urban areas ( OR: 1.53, 95% CI: 1.19-1.97) and being Han ethnicity ( OR: 5.96, 95% CI: 4.47-7.96) were risk factors for exclusive cigarette use, while having a university education or higher was a protective factor ( OR: 0.28, 95% CI: 0.18-0.42). However, for female dual use of cigarettes and cigars, no significant effects were observed for any demographic characteristics. Conclusions:The use rate of cigarettes alone in China is significantly higher than that of cigarette-cigar dual use, and the rates of cigarette use alone and cigarette-cigar dual use in men are significantly higher than those in women. Tobacco use is being affected by sociodemographic factors, among which place of residence, ethnicity and education level are the main influencing factors of cigarette use alone, and gender, age and education level are the main influencing factors of cigarette-cigar dual use.

Objective:To explore the distribution characteristics of glioma-associated oncogene homolog 1 (Gli1) positive cells during orthodontic tooth movement process and conduct a proteomic analysis of these cells.Methods:Forty Gli1-LacZ transgenic mice were used to establish an in vivo orthodontic tooth movement (OTM) model for labeling Gli1 positive cells in Gli1-LacZ transgenic mice (OTM group) and an unforced control group, with tooth movement distance measured using micro-CT. The spatial relationship and distribution characteristics of Gli1 positive cells and H-type vessels of CD31 and endomucin (EMCN) in periodontal tissues were detected by immunofluorescence staining. Twenty Gli1-membrane-targeted tandem dimer Tomato (mT)/membrane-targeted green fluorescent protein (mG) double-genotype mice were bred and Gli1 positive cells were sorted for proteomic sequencing after tamoxifen induction. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases were used for enrichment analysis. Results:The micro-CT three-dimensional reconstruction results showed that the mesial movement of the maxillary first molar in mice after 7 days of force application was (69±15) μm, indicating the successful establishment of the Gli1-LacZ transgenic mouse OTM model. Immunofluorescence staining showed that the blood vessels in periodontal tissue were mostly H-type vessels of CD31 and EMCN. The blood vessels in the periodontal tissues are predominantly H-type vessels positive for both CD31 and EMCN. The percentage of Gli1 positive cells in the OTM group, expressed as (54.5±13.2)%, and the relative fluorescence intensity, expressed as 2.6±0.9, were both significantly greater than those in the control group, which had a Gli1 positive cell percentage of (36.3±9.1)% ( t=3.60 , P=0.002) and a relative fluorescence intensity of 1.0±0.3 ( t=5.20, P<0.001). In contrast to the control group where only a small number of Gli1 positive cells were consistent with the distribution of H-type vessels, in the OTM group the number of Gli1 positive cells increased on the tension side were closely associated with the spatial distribution of H-type vessels. GO enrichment analysis of biological processes found that a large number of proteins in Gli1 positive cells were enriched in pathways such as angiogenesis and tissue remodeling. KEGG enrichment analysis found that related proteins were mainly enriched in pathways related to angiogenesis and Gli1, such as hypoxia-inducing factor 1 signaling pathway, vascular endothelial growth factor signaling pathway and hedgehog signaling pathway. Conclusions:The number of Gli1 positive cells increased on tension side and were closely related to H-type blood vessels in response to mechanical force during orthodontic tooth movement. This may be related to profile of inducing blood vessel formation and tissue remodeling.