Objective:To establish and assess the precision of pre-surgical condyle position planning using mandibular movement trajectory data for orthognathic surgery.Methods:Skull data from large-field cone beam computed tomography(CBCT)and dental oral scan data were imported into IVSPlan 1.0.25 software for 3D reconstruction and fusion,creating 3D models of the maxilla and mandible.Trajectory da-ta of mandibular movement were collected using a mandibular motion recorder,and the data were inte-grated with the jaw models within the software.Subsequently,three-dimensional trajectories of the con-dyle were obtained through matrix transformations,rendering them visually accessible.A senior oral and maxillofacial surgeon with experience in both diagnosis and treatment of temporomandibular joint disease and orthognathic surgery selected the appropriate condyle position using the condyle movement trajectory interface.During surgical design,the mobile mandibular proximal segment was positioned accordingly.Routine orthognathic surgical planning was completed by determining the location of the mandibular distal segment,which was based on occlusal relationships with maxilla and facial aesthetics.A virtual mandible model was created by integrating data from the proximal and distal segment bone.Subsequently,a solid model was generated through rapid prototyping.The titanium plate was pre-shaped on the mandibular model,and the screw hole positions were determined to design a condylar positioning guide device.In accordance with the surgical plan,orthognathic surgery was performed,involving mandibular bilateral sagittal split ramus osteotomy(SSRO).The distal segment of the mandible was correctly aligned inter-maxillary,while the proximal bone segment was positioned using the condylar positioning guide device and the pre-shaped titanium plate.The accuracy of this procedure was assessed in a study involving 10 patients with skeletal class Ⅱ malocclusion.Preoperative condyle location planning and intraoperative po-sitioning were executed using the aforementioned techniques.CBCT data were collected both before the surgery and 2 weeks after the procedure,and the root mean square(RMS)distance between the preope-rative design position and the actual postoperative condyle position was analyzed.Results:The RMS of the condyle surface distance measured was(1.59±0.36)mm(95％CI:1.35-1.70 mm).This value was found to be significantly less than 2 mm threshold recommended by the expert consensus(P＜0.05).Conclusion:The mandibular trajectory may play a guiding role in determining the position of the mandibular proximal segment including the condyle in the orthognathic surgery.Through the use of a con-dylar positioning guide device and pre-shaped titanium plates,the condyle positioning can be personalized and customized with clinically acceptable accuracy.

BACKGROUND:Studies have found that activation of nuclear factor-erythroid 2-related factor 2/heme oxidase-1(Nrf2/HO-1)pathway can alleviate oxidative stress caused by cerebral ischemia-reperfusion injury,but whether human bone marrow mesenchymal stem cells(hBMSC)can activate Nrf2/HO-1 pathway to alleviate cerebral ischemia-reperfusion injury is still lacking relevant studies. OBJECTIVE:To investigate whether intracranial transplantation of hBMSC alleviates oxidative stress injury in cerebral ischemia-reperfusion animal models by activating Nrf2/HO-1 pathway. METHODS:Totally 40 male SPF SD rats were randomly divided into sham operation group,model group,hBMSC transplantation group,hBMSC+solvent group and hBMSC+Nrf2 inhibitor group.Each group consisted of eight animals.In the model group and the hBMSC transplantation group,middle cerebral artery occlusion model was prepared by thread embolization method.The thread embolization was removed 1 hour later,and 30 μL PBS or hBMSC cultured to at least passage 5 was injected into the right cortex and striatum of rats.In the hBMSC+Nrf2 inhibitor group and hBMSC+solvent group,the left ventricle was injected with Nrf2 inhibitor Brusatol and its solvent dimethyl sulfoxide respectively 24 hours before model establishment,then the middle cerebral artery occlusion model was prepared,and hBMSC was injected.Relevant indexes were detected 3 days after transplantation. RESULTS AND CONCLUSION:(1)CT and TTC staining showed the same area and volume of cerebral infarction:model group>hBMSC+Nrf2 inhibitor group>hBMSC+solvent group>hBMSC transplantation group>sham operation group.(2)Hematoxylin-eosin staining and Nissl's staining showed that the ischemic brain tissue was intact and the neurons were normal in the sham operation group.Compared with the model group,the pathological morphology and neuronal injury of the hBMSC transplantation group and the hBMSC+solvent group were significantly improved.Compared with the hBMSC+solvent group,the hBMSC+Nrf2 inhibitor group had more serious pathological morphology and neuronal damage.(3)Western blot assay and oxidative stress index detection results showed that compared with the sham operation group,Nrf2 and HO-1 proteins were decreased(all P<0.05),malondialdehyde was increased and superoxide dismutase was decreased(all P<0.05)in the model group.Compared with the model group,the expression levels of Nrf2 and HO-1 proteins were increased(all P<0.05),malondialdehyde was decreased and superoxide dismutase was increased(all P<0.05)in the hBMSC transplantation group and the hBMSC+solvent group.Compared with the hBMSC+solvent group,the expression levels of Nrf2 and HO-1 proteins were simultaneously decreased(all P<0.05),and malondialdehyde was increased and superoxide dismutase was decreased(all P<0.05)in the hBMSC+Nrf2 inhibitor group.(4)These results indicate that hBMSC can alleviate cerebral ischemia-reperfusion injury possibly by activating Nrf2/HO-1 pathway.

Objective:To observe the effect of metformin on intestinal metabolites and its protective effect on lung injury in an elderly sepsis rat.Methods:SD rats were fed at the Animal Laboratory Center of Zhengzhou University, fourteen elderly SD rats were randomly divided into three groups: sham surgery (age-Sham, AgS group, n=4), cecal ligation and perforation induced sepsis (age-Cecal ligation and puncture, AgCLP group, n=5), and oral administration of metformin (100 mg/kg) after 1 h of CLP treatment (age-Metformin, AgMET group, n=5). Collected rat feces 24 h after modeling, and analyzed the composition and inter group differences of metabolites in the feces using liquid chromatography tandem mass spectrometry non targeted metabolomics. Collected rat lung tissues and detected the expression levels of inflammation related genes and pathological changes in the tissue. The visualization of metabolic changes between groups were presented using orthogonal partial least squares discriminant analysis, heatmaps, and unsupervised principal component analysis, respectively. MetaboAnalyst 3.0 was used to evaluate the Pathway analysis of metabolites, and this software was based on the KEGG database and the human metabolome database. Results:The expressions of CCL4 ( F=203.00, P<0.001), CXCL1( F=65.69, P<0.001), IL-6 ( F=38.94, P<0.002), TNF-α ( F=14.85, P=0.005) between two groups of rats were significantly different (all P<0.05). However, there was no significant difference in CCL2 expression between AgCLP group and AgMET group. Furthermore, compared with the AgS group, the relative intensities of 17 metabolites such as 7-methylxanthine, N-Arachidonylglycine and Manolide in AgCLP group were significantly increased, whereas the 9 metabolites such as Phenazone, Gly-Phe and Valyproline were significantly decreased, and metformin treatment could reverse these changes of the above metabolites. Correlation analysis showed that the IL-6 and TNF-α levels were positively correlated with the relative strength of 7-Methylxanthine, N-Arachidonylglycine and other metabolites, but negatively correlated with the Phenazone and Gly-Phe. CCL4 and CXCL1 were positively correlated with Manolide, but negatively correlated with Valyproline. Conclusion:The results of this study showed that metformin improved sepsis induced acute lung injury and regulates the host intestinal metabolites, which might provide a potential and effective treatment for elderly sepsis induced acute lung injury.

Objective:To explore the mechanism of hepatitis B virus(HBV)inhibiting M1 macrophages to promote immune escape,and to provide targets and strategies for antiviral therapy.Methods:The human monocyte cell line THP-1 was induced into M1 macrophages with PMA+LPS+IFN-γ.Cell morphological changes and the expressions of CD68,CD86,HLA-DR and functional molecules IL-1β,IL-6,TNF-α in M1 macrophages were detected by flow cytometry and RT-qPCR to identify M1 macrophages.HBV stable replication cell line HepG2.2.15 were co-cultured with M1 macrophages,and the expression of HBV-DNA was detected by qP-CR.The expression of CD68,CD86 and HLA-DR were detected by flow cytometry.The expressions of functional molecules TLR4,NLRP3,Caspase-1,pro-caspase-1,caspase-1 p20,IL-1β and IL-18 in M1 macrophages were determined by RT-qPCR and Western blot.Apoptosis rate was detected by flow cytometry,and the expression of apoptosis related protein cleaved-caspase-3 was determined by Western blot.Results:THP-1 was successfully induced to differentiate into M1 macrophages.M1 macrophages inhibited HBV repli-cation(P<0.05).HBV inhibited the expressions of CD68,CD86 and HLA-DR in M1 macrophages(P<0.01).HBV inhibited the ex-pressions of TLR4,NLRP3,Caspase-1,caspase-1 p20,IL-1β and IL-18 in M1 macrophages(P<0.01).HBV induced M1 macro-phage apoptosis(P<0.05).Conclusion:HBV inhibits M1 macrophages and their functional molecules TLR4,NLRP3 and down-stream factors,reduces the synthesis and secretion of inflammatory factors,induces apoptosis,and then promotes immune escape,re-sulting in the persistence and replication of HBV in the body.

BACKGROUND: Microsatellite instability (MSI) is a key biomarker for cancer immunotherapy and prognosis. Integration of MSI testing into a next-generation-sequencing (NGS) panel could save tissue sample, reduce turn-around time and cost, and provide MSI status and comprehensive genomic profiling in single test. We aimed to develop an MSI calling model to detect MSI status along with the NGS panel-based profiling test using tumor-only samples. METHODS: From January 2019 to December 2020, a total of 174 colorectal cancer (CRC) patients were enrolled, including 31 MSI-high (MSI-H) and 143 microsatellite stability (MSS) cases. Among them, 56 paired tumor and normal samples (10 MSI-H and 46 MSS) were used for modeling, and another 118 tumor-only samples were used for validation. MSI polymerase chain reaction (MSI-PCR) was performed as the gold standard. A baseline was built for the selected microsatellite loci using the NGS data of 56 normal blood samples. An MSI detection model was constructed by analyzing the NGS data of tissue samples. The performance of the model was compared with the results of MSI-PCR. RESULTS: We first intersected the target genomic regions of the NGS panels used in this study to select common microsatellite loci. A total of 42 loci including 23 mononucleotide repeat sites and 19 longer repeat sites were candidates for modeling. As mononucleotide repeat sites are more sensitive and specific for detecting MSI status than sites with longer length motif and the mononucleotide repeat sites performed even better than the total sites, a model containing 23 mononucleotide repeat sites was constructed and named Colorectal Cancer Microsatellite Instability test (CRC-MSI). The model achieved 100% sensitivity and 100% specificity when compared with MSI-PCR in both training and validation sets. Furthermore, the CRC-MSI model was robust with the tumor content as low as 6%. In addition, 8 out of 10 MSI-H samples showed alternations in the four mismatch repair genes ( MLH1 , MSH2 , MSH6 , and PMS2 ). CONCLUSION MSI status can be accurately determined along the targeted NGS panels using only tumor samples. The performance of mononucleotide repeat sites surpasses loci with longer repeat motif in MSI calling.
Humans ; Microsatellite Instability ; Colorectal Neoplasms/diagnosis* ; Microsatellite Repeats/genetics* ; DNA Mismatch Repair

Objectives:To analysis the effect of continuous positive airway pressure (CPAP) on nocturnal blood pressure in patients complicated with obstructive sleep apnea-hypopnea syndrome (OSAHS) and different circadian rhythms of hypertension.Methods:A total of 61 eligible patients were monitored by overnight polysomnography (PSG) at the Sleep Center of the Affiliated Huaian No.1 People′s Hospital of Nanjing Medical University between January 2020 and April 2021. During the period of PSG monitoring, continuous non-invasive blood pressure (BP) and heart rate variability (HRV) were monitored simultaneously. Frequency domain analysis was used to measure HRV and low/high frequency was used to indirectly reflect sympathetic activity. According to the nighttime systolic BP decrease rate, patients were divided into three groups: dipper pattern (descent rate ≥10%), non-dipper pattern (descent rate was less than 10% but higher than 0) and reverse dipper pattern (descent rate≤0). The PSG parameters, BP data as well as sympathetic activity etc. were compared within and among groups before and after CPAP treatment. Multiple linear regression analyses were used to explore the influencing factors of antihypertensive effect of CPAP.Results:There were no significant differences in awake systolic BP (SBP) values, the severity of OSAHS, ESS scores, awake sympathetic activity and the other baseline data among the three groups. After CPAP treatment, the mean value of asleep BP in entire group showed a modest decline as compared to the baseline values [SBP decreased 4.6 mmHg (1 mmHg=0.133 kPa); diastolic blood pressure (DBP) decreased 2.4 mmHg, both P<0.001]. Subgroup analysis showed a significant reduction in asleep SBP of 11.1 mmHg and DBP of 4.9 mmHg (both P<0.001) in reverse dipper group, respectively, compared with the baseline values. While in dipper and non-dipper group, there were no significant differences before and after CPAP treatment in terms of BP (both P>0.05). In addition, there was no difference in awake sympathetic activity among three groups, while sleep sympathetic activity showed a gradual increasing trend. Sleep sympathetic activity decreased significantly from baseline after CPAP treatment in reverse dipper group ( P<0.001), while no differences were found in the other two groups before and after treatment. After controlling for baseline data such as age etc., the line regression model showed that the antihypertensive effect of CPAP was correlated with reverse dipper (SBP: β=0.548, P=0.002; DBP: β=0.454, P=0.013) and the improvement of nocturnal MpO 2 (SBP: β=0.410, P=0.046), but not with the severity of OSAHS, daytime sleepiness, or baseline BP values. Conclusion:For patients with moderate to severe OSAHS and hypertension, reverse dipper is an effective indicator to predict the antihypertensive effect of CPAP therapy.

Senescence of activated hepatic stellate cells (aHSCs) is a stable growth arrest that is implicated in liver fibrosis regression. Senescent cells often accompanied by a multi-faceted senescence-associated secretory phenotype (SASP). But little is known about how alanine-serine-cysteine transporter type-2 (ASCT2), a high affinity glutamine transporter, affects HSC senescence and SASP during liver fibrosis. Here, we identified ASCT2 is mainly elevated in aHSCs and positively correlated with liver fibrosis in human and mouse fibrotic livers. We first discovered ASCT2 inhibition induced HSCs to senescence in vitro and in vivo. The proinflammatory SASP were restricted by ASCT2 inhibition at senescence initiation to prevent paracrine migration. Mechanically, ASCT2 was a direct target of glutaminolysis-dependent proinflammatory SASP, interfering IL-1α/NF-κB feedback loop via interacting with precursor IL-1α at Lys82. From a translational perspective, atractylenolide III is identified as ASCT2 inhibitor through directly bound to Asn230 of ASCT2. The presence of -OH group in atractylenolide III is suggested to be favorable for the inhibition of ASCT2. Importantly, atractylenolide III could be utilized to treat liver fibrosis mice. Taken together, ASCT2 controlled HSC senescence while modifying the proinflammatory SASP. Targeting ASCT2 by atractylenolide III could be a therapeutic candidate for liver fibrosis.

Objectives:To investigate the common risk factors for excess daytime sleepiness (EDS) and hypertension in obstructive sleep apnea-hypopnea syndrome(OSAHS) patients.Methods:Between January 2020 and February 2021, a total of 103 OSAHS patients diagnosed in the Department of Sleep Medicine Center, the Affiliated Huaian No.1 People′s Hospital of Nanjing Medical University were enrolled as the study population. During polysomnography (PSG) monitoring, noninvasive continuous blood pressure (BP) and heart rate variability (HRV) were monitored simultaneously. Low/high frequency components (LF/HF) were used to reflect sympathetic-vagal balance in frequency domain analysis. According to Epworth Sleepiness Scale (ESS) and BP levels, patients were divided into four groups: simple OSAHS group (ESS<10 scores and BP<140/90 mmHg, n=30)(1 mmHg=0.133 kPa), OSAHS+hypertension group (ESS<10 scores and BP≥140/90 mmHg, n=23), OSAHS+EDS group (ESS≥10 scores and BP<140/90 mmHg, n=26) and OSAHS+hypertension+EDS group (ESS≥10scores and BP≥140/90 mmHg, n=24). The clinical and PSG parameters were analyzed and compared among the four groups. Regression analyses were used to explore the common causative factors for EDS and hypertension. Results:The LF/HF in OSAHS+hypertension+EDS group was significantly higher than the other three groups [3.2% (2.6%, 4.2%) vs 1.4% (1.2%, 1.6%), 2.2% (1.8%, 2.9%), 2.5% (1.6%, 3.1%), all P<0.05]. No difference was observed between OSAHS+hypertension group and OSAHS+EDS group ( P=0.779), but both higher than simple OSAHS group. The linear regression equation showed that LF/HF was most correlated with the percentage of sleep time with oxygen saturation<90% (T90) as compared to the other parameters of sleep disordered breathing (β=0.201, P=0.006). In addition, Pearson correlation analysis showed that LF/HF was significantly correlated with ESS scores and asleep BP levels ( r=0.536, r=0.456, all P<0.05). The logical regression equation showed that LF/HF was a causative risk factor for both EDS and hypertension in OSAHS (β=0.164, 95% CI: 1.018-1.364, P=0.028). Conclusion:The sympathetic-vagal imbalance is a common risk factor for EDS and hypertension in OSAHS patients

Objective:To investigate the association between oxygen desaturation rate and blood pressure (BP) among severe obstructive sleep apnea syndrome (OSAS) and the possible mechanism.Methods:Patients with snoring were enrolled from the Department of Sleep Medicine Center, the Affiliated Huaian No.1 People′s Hospital of Nanjing Medical University form March 2018 to January 2019 and underwent polysomnography (PSG). Noninvasive BP and Heart rate variability were full-night monitored continuously and synchronized with PSG. Based on the PSG results and exclusion criteria, a total of 86 severe OSAS patients were enrolled in this study and divided into two groups according to the ambulatory BP measurements: hypertensive group ( n=44) and normotensive group ( n=42). Oxygen desaturation rate was expressed as the change in the percentage of pulse oxyhemoglobin saturation (SpO 2) per second during desaturation events after the obstructive apnea events occurred. The PSG parameters were compared between the two group and the multiple regression analyses were used to explore the association between oxygen desaturation rate and BP and its possible mechanism. Results:The apnea-hyperpnoea index (AHI) and respiratory event-related arousals (RERAs) were significantly higher in hypertensive group than those in normotensive group [(69.8±18.2) vs. (56.5±13.9) event/h; (40.5±17.4) vs. (30.2±14.6) event/h, both P<0.01]. In addition, hypoxia exposure conditions in the hypertensive group were more severe than those in the normotensive group, especially oxygen desaturation rate [(0.45±0.14)%/s vs. (0.33±0.10)%/s, P<0.001]. After adjusting for age, sex, neck circumference, waist circumference, smoking, drinking, the regression analyses showed that only the oxygen desaturation rate was significantly associated with both awake and asleep BP in OSAS patients ( β=0.473, 0.478, both P<0.01) and the correlation analyses suggested that the oxygen desaturation rate was related to the both awake and asleep sympathetic-parasympathetic imbalance ( r=0.367, 0.337, both P<0.01). Conclusion:Oxygen desaturation rate is closely related to BP levels in patients with severe OSAS, and the underlying mechanism is associated with the increased sympathetic activity.

Objectives To determine the risk factors of ventilator-associated pneumonia (VAP) in the pediatric intensive care unit and to explore effective strategies to reduce the morbidity of VAP. Methods A retrospective analysis was conducted on 455 children admitted into the PICU of Hunan Children's Hospital from June 2014 to June 2017. The 455 children were divided into VAP group (n=43) and non-VAP group (n=412). The incidence of VAP was identified and risk factors were compared using the logistic regression analysis via SPSS 19.0 software.Results There were 311 males and 144 females with a median age of 11 months old (29 days to 9 years and 4 months). The incidence of VAP was 9.45% (43/455). Congenital laryngeal and trachea malformation with pulmonary infection was the first reason for the occurrence of VAP (23.3%), followed by congenital heart diseases with pulmonary infection (18.6%). Via univariate analysis, types of endotracheal intubation (χ2=45.33, P<0.001), duration of mechanical ventilation (Z=1.21, P=0.034), re-intubation (χ2=20.22, P=0.004), early usage of antibiotics (χ2=4.98, P=0.026),and methods of nutritional support(χ2=10.15,P=0.006)were identified as risk factors of VAP in the pediatric intensive care unit patients (P<0.05). Based on the multivariate logistic regression analysis, the followings were all independent predictor for VAP:types of endotracheal intubation(OR=1.87,95%CI:1.48~9.75),duration of mechanical ventilation(OR=1.14, 95%CI:1.08~2.35), re-intubation (OR=3.42, 95%CI:1.26~5.57), early usage of antibiotics (OR=4.55, 95%CI:2.21~8.77). Conclusions Many risk factors were found related with the occurrence of VAP. A comprehensive analysis of the host factors and iatrogenic factors should be conducted. Rational use of antibiotics and daily assessment of extubation might help reduce the incidence of VAP.