ObjectiveTo establish a qualitative and quantitative analysis method for chemical constituents in Liu Junzitang（LJZT）， and to clarify its material basis. MethodThe chemical constituents in LJZT were analyzed by ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS）， and the resulting compounds were identified by using databases， such as MassBank， PubChem， ChemSpider， Traditional Chinese Medicine Systems Pharmacology Database and Analytical Platform（TCMSP）， and by combining with relevant literature. UPLC was used to establish a quantitative method for analysis of 9 compounds in LJZT， including liquiritin， hesperidin， lobetyolin， liquiritigenin， glycyrrhizic acid， nobiletin， tangeretin， atractylenolide Ⅱ and Ⅰ. ResultBy combining the relevant literature， database and MS information， a total of 79 compounds were identified from LJZT， including 31 flavonoids， 15 terpenoids， 14 nitrogen-containing compounds， 6 phenylpropanoids， 6 organic acids and 7 other compounds. The established quantitative analytical method for the nine representative components showed good linearity within their respective linear ranges， and the precision， stability， reproducibility and recovery were in accordance with the requirements. The quantitative results showed that the contents of liquiritin， hesperidin， lobetyolin， liquiritigenin， glycyrrhizic acid， nobiletin， tangeretin， atractylenolide Ⅱ and Ⅰ in LJZT were 0.376 5， 2.602 1， 0.082 6， 0.128 1， 1.778 6， 0.015 7， 0.006 7， 0.030 4， 0.003 2 mg·g^-1， respectively. ConclusionThe established method can quickly， sensitively and accurately analyze the chemical constituents in LJZT， clarify that the material basis of LJZT is mainly flavonoids， terpenoids and nitrogen-containing compounds， and simultaneously determine the contents of the 9 components， which can lay a foundation for the research on quality control， mechanism and clinical application of LJZT.

Objective To observe the effect of a new cell delivery tool(MSC exo)on the proliferation of pancreatic cancer by transferring targeted genes.Methods Transmission Electron Microscope(TEM)and Nanoparticle Tracking Analysis(NTA)were used to identify human mesenchymal stem cell exosomes(MSC-exo)and transport miR-450a-5p into CFPAC-1,to explore the effect of miR-450a-5p targeting BZW2 on inhibiting the proliferation of pancreatic cancer cells.Results The expression of miR-450a-5p was low in pancreatic cancer tissue(P<0.05),and the expression of CD63 and TSG101 of MSC-exo-miR-450a-5p in CFPAC-1 cells was higher than that of MSC-exo by Western blot(P<0.05).CCK-8 and EdU results showed that MSC-exo-miR-450a-5p significantly inhibited the proliferation of CFPAC-1 cells(P<0.05).Cell scratch and Transwell experiments showed that MSC-exo-miR-450a-5p can inhibit the migration and invasion of CFPAC-1 cells(P<0.05).Through dual luciferase assay,it was confirmed that miR-450a-5p targets BZW2,and RT-qPCR and Western blotting showed a negative correlation(P<0.05)between miR-450a-5p and BZW2 expression.Overexpression of BZW2,CCK-8,EdU,cell scratch,and Transwell experiments confirmed that pc-BZW2 reversed the anti-cancer function of MSC-exo-miR-450a-5p on CFPAC-1.Western blot detected PCNA,Ki-67,MMP2,MMP9,and the results were consistent with the above experiments(P<0.05).Conclusion hMSC exo is a new delivery system,targeting BZW2 to transport miR-450a-5p to inhibit the biological malignancy of pancreatic cancer cells,which provides an important clue for the research of targeted treatment of pancreatic cancer.

Objective:To evaluate the relationship between hippocampal receptor-interacting protein kinase-1 (RIPK1) and nucleotide-binding oligomerization domain-like receptor containing pyrin domain 3 (NLRP3) inflammasomes in postoperative neurocognitive dysfunction of aged rats with chronic knee arthritis pain.Methods:Sixty-four healthy male Sprague-Dawley rats, aged 18 months, weighing 500-550 g, were divided into 4 groups ( n=16 each) using a random number table method: chronic knee arthritis pain group (group P), chronic knee arthritis pain+ operation group (group PS), RIPK1 inhibitor necrostatin-1+ chronic knee arthritis pain+ operation group (group NPS), and DMSO+ chronic knee arthritis pain+ operation group (group DPS). The knee arthritis model was prepared by intra-articular injection of monosodium iodoacetate (MIA) 1 mg into the left knee joint, and 12 weeks later exploratory laparotomy was performed under sevoflurane anesthesia. Necrostatin-1 6.25 mg/kg and the equal volume of DMSO were intraperitoneally injected at 1 h before operation in NPS group and DPS group, respectively. Thermal pain threshold was measured at 1 week before MIA injection and 6 and 12 weeks after MIA injection. Morris water maze test was used to evaluate the cognitive function at 7 days after surgery. Hippocampal tissues were obtained for microscopic examination of the pathological changes (after HE staining) and for determination of the expression of RIPK1, phosphorylated RIPK1 (p-RIPK1), NLRP3, activated cysteine-aspartic protease caspase-1 (cl-caspase-1), apoptosis-associated speck-like protein containing a CARD (ASC) (by Western blot) and contents of interleukin-1beta (IL-1β) and IL-18 (by enzyme-linked immunosorbent assay). Results:Thermal pain threshold was significantly decreased at 6 and 12 weeks after MIA injection as compared with that before injection ( P<0.05), and there was no significant difference in thermal pain threshold among the four groups ( P>0.05). Compared with P group, the escape latency was significantly prolonged, the time of staying at the original platform quadrant was shortened, the number of crossing the original platform was reduced, the expression of RIPK1, p-RIPK1, NLRP3, cl-caspase-1 and ASC was up-regulated, and the contents of IL-1β and IL-18 were increased ( P<0.05), and pathological changes of hippocampal neurons were marked in PS group, DPS group and NPS group. Compared with PS group and DPS group, the escape latency was significantly shortened, the time of staying at the original platform quadrant was prolonged, the number of crossing the original platform was increased, the expression of RIPK1, p-RIPK1, NLRP3, cl-caspase-1 and ASC was down-regulated, the contents of IL-1β and IL-18 were decreased ( P<0.05), and pathological changes of hippocampal neurons were significantly attenuated in NPS group. Conclusions:Postoperative hippocampal RIPK1 function is enhanced in aged rats with chronic knee arthritis pain, which then activates NLRP3 inflammasomes, triggering neuroinflammation, and this process may be involved in the mechanism of postoperative neurocognitive dysfunction.

Malocclusion,identified by the World Health Organization(WHO)as one of three major oral diseases,profoundly impacts the dental-maxillofacial functions,facial esthetics,and long-term development of～260 million children in China.Beyond its physical manifestations,malocclusion also significantly influences the psycho-social well-being of these children.Timely intervention in malocclusion can foster an environment conducive to dental-maxillofacial development and substantially decrease the incidence of malocclusion or reduce the severity and complexity of malocclusion in the permanent dentition,by mitigating the negative impact of abnormal environmental influences on the growth.Early orthodontic treatment encompasses accurate identification and treatment of dental and maxillofacial morphological and functional abnormalities during various stages of dental-maxillofacial development,ranging from fetal stages to the early permanent dentition phase.From an economic and societal standpoint,the urgency for effective early orthodontic treatments for malocclusions in childhood cannot be overstated,underlining its profound practical and social importance.This consensus paper discusses the characteristics and the detrimental effects of malocclusion in children,emphasizing critical need for early treatment.It elaborates on corresponding core principles and fundamental approaches in early orthodontics,proposing comprehensive guidance for preventive and interceptive orthodontic treatment,serving as a reference for clinicians engaged in early orthodontic treatment.

Background Public places are frequently polluted by cigarette smoking, and there is a lack of accurate, real-time, and intelligent monitoring technology to identify smoking behavior. It is necessary to develop a tool to identify cigarette smoking behavior in public places for more efficient control of cigarette smoking and better indoor air quality. Objective To construct a model for recognizing cigarette smoking behavior based on real-time indoor concentrations of PM_2.5 in public places. Methods Real-time indoor PM_2.5 concentrations were measured for at least 7 continuous days in 10 arbitrarily selected places (6 public service providers and and 4 office or other places) from Oct. to Nov. 2022 in Pudong New Area, Shanghai. Indoor nicotine concentrations were monitored with passive samplers simultaneously. Outdoor PM_2.5 concentration data were obtained from three municipal environmental monitoring stations which were nearest to each monitoring point during the same period. Mann-Whitney U test was used to compare indoor and outdoor means of PM_2.5 concentrations, and Spearman rank correlation was used to analyze indoor PM_2.5 and nicotine concentrations. An interactive plot and a random forest model was applied to examine the association between video observation validated indoor smoking behavior and real-time indoor PM_2.5 concentrations in an Internet cafe. Results The average indoor PM_2.5 concentration in the places providing public services [(97.5±149.3) µg·m⁻³] was significantly higher than that in office and other places [(19.8±12.2) µg·m⁻³] (P=0.011). The indoor/outdoor ratio (I/O ratio) of PM_2.5 concentration in the public service providers ranged from 1.1 to 19.0. Furthermore, the indoor PM_2.5 concentrations in the 10 public places were significantly correlated with the nicotine concentrations (r_s=0.969, P<0.001). Among them, the top 3 highly polluted places were Internet cafes, chess and card rooms, and KTV. The results of random forest modeling showed that, for synchronous real-time PM_2.5 concentration, the area under the curve (AUC) was 0.66, while for PM_2.5 concentration at a lag of 4 min after the incidence of smoking behavior, the AUC increased to 0.72. Conclusion The indoor PM_2.5 concentrations in public places are highly correlated with smoking behavior. Based on real-time indoor PM_2.5 monitoring, a preliminary recognition model for smoking behavior is constructed with acceptable accuracy, indicating its potential values applied in smoking control and management in public places.

Objective:To investigate the effect of homocysteine (Hcy) on cerebral perfusion and cognitive function in patients with arteriosclerotic cerebral small vessel disease (aCSVD).Methods:A total of 117 patients with aCSVD who visited the First Affiliated Hospital of Anhui Medical University from June 2020 to September 2022 were enrolled and divided into the aCSVD cognitive impairment group (aCSVD-CI, n=57) and aCSVD non-cognitive impairment group (aCSVD-NCI, n=60) according to the Montreal Cognitive Assessment score. Serum Hcy measurement, cognitive function assessment, and three-dimensional pseudo-continuous arterial spin labeling perfusion imaging scan were performed in all patients, and multivariate Logistic regression analysis was used to explore risk factors for cognitive impairment in patients with aCSVD. The cerebral blood flow and perfusion differential brain regions of the whole brain, grey matter, and white matter were compared between the two groups. Partial correlation analyses were performed between the serum Hcy, overall cognitive function scores and cerebral blood flow in grey matter, as well as between the cerebral blood flow in the perfusion differential brain area and cognitive function scores. The mediating effect model was used to analyze the role of grey matter blood flow in the relationship between serum Hcy and overall cognition. Results:The serum Hcy level in the CSVD-CI group was higher than that in the CSVD-NCI group [16.38(14.02, 18.58) μmol/L vs 14.40 (11.93, 15.73) μmol/L, Z=-3.81, P<0.001]. In terms of cerebral perfusion, compared with the aCSVD-NCI group, the aCSVD-CI group had significantly lower cerebral blood flow in grey matter ( Z=-3.22, P=0.001), left middle frontal gyrus ( t=-4.91, P<0.05), right middle frontal gyrus ( t=-5.14, P<0.05), and right orbital medial frontal lobe ( t=-4.38, P<0.05). In contrast, the left hippocampus ( t=4.58, P<0.05) had increased cerebral blood flow. Multivariate Logistic regression analysis showed that serum Hcy level was independent risk factor for cognitive impairment in aCSVD after controlling for multiple risk factors. Partial correlation analysis showed that left middle frontal gyrus blood flow ( r=-0.39, P=0.006), right middle frontal gyrus blood flow ( r=-0.44, P=0.002), and right orbital medial frontal lobe cerebral blood flow ( r=-0.43, P=0.002) were negatively correlated with the Stroop Color Word Test-C results. Left hippocampal cerebral blood flow was negatively correlated with Auditory Word Learning Test-long-delayed recall ( r=-0.43, P=0.002). Further mediation analysis showed that the effect of Hcy on cognitive function was partly mediated by grey matter cerebral blood flow (indirect effect=-0.11, P<0.001). Conclusion:Hcy is an independent risk factor for cognitive impairment in aCSVD, and part of the effect of elevated Hcy on cognitive impairment in aCSVD may be mediated by decreased gray matter cerebral perfusion.

Orthodontically induced tooth root resorption (OIRR) is a serious complication during orthodontic treatment. Stimulating cementum repair is the fundamental approach for the treatment of OIRR. Parathyroid hormone (PTH) might be a potential therapeutic agent for OIRR, but its effects still lack direct evidence, and the underlying mechanisms remain unclear. This study aims to explore the potential involvement of long noncoding RNAs (lncRNAs) in mediating the anabolic effects of intermittent PTH and contributing to cementum repair, as identifying lncRNA-disease associations can provide valuable insights for disease diagnosis and treatment. Here, we showed that intermittent PTH regulates cell proliferation and mineralization in immortalized murine cementoblast OCCM-30 via the regulation of the Wnt pathway. In vivo, daily administration of PTH is sufficient to accelerate root regeneration by locally inhibiting Wnt/β-catenin signaling. Through RNA microarray analysis, lncRNA LITTIP (LGR6 intergenic transcript under intermittent PTH) is identified as a key regulator of cementogenesis under intermittent PTH. Chromatin isolation by RNA purification (ChIRP) and RNA immunoprecipitation (RIP) assays revealed that LITTIP binds to mRNA of leucine-rich repeat-containing G-protein coupled receptor 6 (LGR6) and heterogeneous nuclear ribonucleoprotein K (HnRNPK) protein. Further co-transfection experiments confirmed that LITTIP plays a structural role in the formation of the LITTIP/Lgr6/HnRNPK complex. Moreover, LITTIP is able to promote the expression of LGR6 via the RNA-binding protein HnRNPK. Collectively, our results indicate that the intermittent PTH administration accelerates root regeneration via inhibiting Wnt pathway. The lncRNA LITTIP is identified to negatively regulate cementogenesis, which activates Wnt/β-catenin signaling via high expression of LGR6 promoted by HnRNPK.
Mice ; Animals ; Cementogenesis ; Wnt Signaling Pathway ; beta Catenin/metabolism* ; Heterogeneous-Nuclear Ribonucleoprotein K/metabolism* ; RNA, Long Noncoding/genetics* ; Parathyroid Hormone ; Receptors, G-Protein-Coupled/metabolism*

Objective:To investigate the global and local changes of neurovascular coupling in arteriosclerotic cerebral small vessel disease (aCSVD) and the correlation with cognitive function.Methods:Forty-three patients with confirmed aCSVD from the outpatient department or ward of the Department of Neurology, the First Affiliated Hospital of Anhui Medical University between June 2020 and June 2021 were enrolled in this study. Meanwhile, 48 healthy subjects were selected as controls. Cognitive evaluation, resting-state functional magnetic resonance imaging and 3D pseudo-continuous arterial spin labeling magnetic resonance imaging scanning were performed in all subjects. The global cerebral blood flow-regional homogeneity (ReHo) correlation coefficient and the cerebral blood flow/ReHo ratio were used to evaluate global and local neurovascular coupling. Meanwhile, correlations between the cerebral blood flow/ReHo ratio and neuropsychological assessments were explored in aCSVD patients.Results:Global cerebral blood flow-ReHo coupling was decreased in aCSVD patients compared to healthy controls [aCSVD patients: 0.942(0.933, 0.950), healthy controls: 0.947(0.939, 0.954), Z=-2.11, P=0.035]. aCSVD patients showed decreased cerebral blood flow/ReHo ratio in the right lingual gyrus ( t=-4.45, P<0.05) and increased cerebral blood flow/ReHo ratio in the left ( t=4.91, P<0.05) and right ( t=4.72, P<0.05) inferior parietal lobule. Cerebral blood flow/ReHo ratio of the right inferior parietal lobule was negatively correlated with total score ( r=-0.33, P=0.031) and praxis score ( r=-0.43, P=0.004) in Cambridge Cognitive Examination-Chinese Version subitems and positively correlated with scores of Stroop Color Word Test (SCWT)-color ( r=0.33, P=0.032), SCWT-word ( r=0.34, P=0.025) and Trail Making Test-B ( r=0.31, P=0.043) in aCSVD patients. While the cerebral blood flow/ReHo ratio of the right lingual gyrus was negatively correlated with Visual Replicate-Immediate Recall score ( r=-0.36, P=0.017). Conclusion:aCSVD patients showed abnormal global and local neurovascular coupling, which was associated with attention, executive function, and visual space function.

Objective:To investigate the predictive value of visceral adipose tissue (VAT) in occult peritoneal metastasis (OPM) of gastric cancer.Methods:A total of 93 patients with gastric cancer admitted to the First Affiliated Hospital of Zhengzhou University from October 2018 to October 2021 were retrospectively collected. None of the patients had typical peritoneal metastasis on CT. Patients were divided into OPM group (31 cases) and non-OPM group (62 cases) according to laparoscopic exploration. The clinical, pathological and CT features were recorded. The parameters related to adipose tissue (VAT and subcutaneous adipose tissue) within the range of 15 mm and 25 mm below the largest layer of gastric cancer lesions in preoperative CT images were measured, including the volume, average CT attenuation and standard deviation. The independent-sample t test, Wilcoxon rank-sum test, χ 2 test or Fisher′s exact probability were used to compare the clinical, pathological and CT imaging parameters between OPM and non-OPM groups. Multivariate logistic regression analysis was used to explore the independent risk factors for OPM of gastric cancer and establish a combined model. The receiver operating characteristic curve was used to evaluate the efficacy of each indicator and the combined model in predicting OPM of gastric cancer. Results:There were statistically significant differences in age, pathological type, CA125, ascites, cT stage, the thickest diameter of lesion, average CT attenuation of 15 mm VAT and 25 mm VAT between the OPM group and the non-OPM group ( P<0.05). Multivariate analysis showed that ascites, cT stage and average CT attenuation of 25 mm VAT were independent risk factors for OPM of gastric cancer, with the OR (95%CI) of 4.940 (1.287-18.967), 4.284 (1.270-14.455), and 1.149 (1.013-1.303), respectively. A combined model was established. The area under the curve (AUC) of ascites, cT stage, average CT attenuation of 25 mm VAT, average CT attenuation of 15 mm VAT and combined model were 0.685, 0.718, 0.703, 0.674 and 0.813, respectively. There were statistically significant differences in AUC between the combined model and each four single indicators above ( Z=2.98, 2.63, 2.09, 2.54, P=0.003, 0.009, 0.037, 0.011). Conclusions:The ascites, cT stage and average CT attenuation of 25 mm VAT are independent risk factors for OPM in gastric cancer. The combined model based on the above three indicators has the best performance in predicting OPM in gastric cancer.

Objective:To evaluate the effect of oral iodized oil pills on iodine nutrition and thyroid function of pregnant women in Xinjiang Uygur Autonomous Region (Xinjiang).Methods:From May to July 2017, one county was selected from Ili Prefecture without taking iodized oil pills, Aksu Prefecture taking iodized oil pills once a year, and Kashgar Prefecture taking iodized oil pills twice a year in Xinjiang as survey sites, respectively; 100 pregnant women (evenly distributed in early, middle and late pregnancy) were selected from each survey county, the general data, urine and blood samples were collected, and urinary iodine and thyroid function indicators [free triiodothyronine (FT 3), free thyroxine (FT 4), thyrotropin (TSH), anti thyroglobulin antibody (TgAb), and anti thyroid peroxidase antibody (TPOAb)] were tested. Results:A total of 308 pregnant women were investigated, and 289 were finally included in the analysis, with an average age of 25 years; the body mass index (BMI) was (22.69 ± 3.07) kg/m 2. The occupation distribution was mainly farmer, accounting for 93.77% (271/289); most of them had junior high school education or below, accounting for 71.97% (208/289). The median urinary iodine of early, middle and late pregnant women in Kashgar Prefecture was 712.87, 604.50 and 656.23 μg/L, respectively, which were in iodine excess state. The iodine nutrition level of early pregnant women in Ili Prefecture and Aksu Prefecture was in iodine super suitability state, and the iodine nutrition level of middle and late pregnant women was in the iodine suitability state. The difference of median urinary iodine in early, middle and late pregnancy women between different regions was statistically significant ( Z = 53.02, 49.60, 44.66, P < 0.001). In addition, the urinary iodine of women in Kashgar Prefecture during each pregnancy period was significantly higher than that in Ili Prefecture and Aksu Prefecture ( P < 0.05). There were statistically significant differences in the levels of FT 3 among women in early pregnancy, FT 4 and TSH among women in middle pregnancy between different regions ( F = 4.59, 10.92, Z = 8.61, P < 0.05 or < 0.001). Among them, the level of FT 3 in early pregnancy in Kashgar Prefecture was lower than that in Ili Prefecture ( P < 0.05); the level of FT 4 in Kashgar Prefecture during middle pregnancy was higher than that in Ili Prefecture, and the level of TSH was lower than that in Ili Prefecture ( P < 0.05). There was no significant difference in TgAb positive rate, TPOAb positive rate and double antibody positive rate of early, middle and late pregnant women between different regions ( P > 0.05). The detection rates of hypothyroxinemia in early pregnant women in Ili Prefecture, Aksu Prefecture and Kashgar Prefecture were 13.9% (5/36), 3.2% (1/31) and 0 (0/33), respectively, and there was a statistically significant difference between different regions ( P = 0.036). The detection rates of subclinical hyperthyroidism in middle pregnant women were 0 (0/35), 0 (0/40), 17.6% (6/34), respectively, and there was a statistically significant difference between different regions ( P = 0.001). Conclusions:The results of urinary iodine in the three regions are in line with the iodine nutrition distribution under their respective iodine supplement strategies. Pregnant women in Kashgar Prefecture present iodine excess status after taking iodized oil pills; at the same time, the serum FT 3, FT 4 and TSH levels of pregnant women in Kashgar Prefecture are affected by iodine nutrition levels. Although it is scientific and effective to implement the intensified iodine supplement measures for pregnant women, it is still necessary to further study the suitability of oral iodine oil pills to ensure that pregnant women are at an suitability iodine nutrition level.