OBJECTIVE To provide a reference for optimizing and promoting the quality standards of Shechuan naolitong granules. METHODS Fifteen batches of Shechuan naolitong granules were used as samples to establish HPLC fingerprints using the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine （2012 edition）. Similarity evaluation and common peak identification were performed， and orthogonal partial least squares discriminant analysis （OPLS-DA） was used to assess quality differences among different batches and to screen quality differential components. Using salvianolic acid B（SAB） as the internal reference， quantitative analysis of multi-components by single-marker （QAMS） was developed to simultaneously determine geniposidic acid （GA）， chlorogenic acid （CA）， vaccarin （VA）， ferulic acid （FA） and senkyunolide I （SI）. The results were compared with those obtained by the external standard method. RESULTS A total of 13 common peaks were identified in the HPLC fingerprints of 15 batches of samples， and the similarities of the spectra were all above 0.96. Seven chromatographic peaks were identified as GA （peak 3）， CA （peak 6）， VA （peak 8）， FA （peak 9）， SI （peak 11）， SAB（peak 12） and TA（peak 13）. OPLS-DA indicated that the differential quality markers among 15 batches were peaks 5， 11 （SI）， and 12 （SAB）.Using SAB as the internal reference， the relative correction factors for GA， CA， VA， FA and SI were calculated as 1.058 4， 0.594 3， 0.643 3， 0.342 7 and 0.262 8， respectively. The mean content of GA， CA， VA， FA， SI and SAB across the 15 batches of samples were 0.155 0， 0.085 4， 0.140 3， 0.071 8， 0.072 7， 1.276 3 mg/g， respectively， showing no significant difference compared with the ESM （P＞0.05）. CONCLUSIONS The established HPLC fingerprint and QAMS are simple， efficient and economical， providing a reference for the quality control and further development of Shechuan naolitong granules.

OBJECTIVES: Digitally collect data on the preparation space of the molar occlusal surface for full crown restoration and analyze the minimum preparation spaces and areas prone to insufficient preparation. METHODS: A total of 846 molars (excluding third molars) designated for repair with zirconium dioxide crowns were selected. All molars were prepared by using the fixed-depth groove method under visual guidance. A digital impression was collected, and the occlusal surface of the preparation was divided into regions: mesiobuccal area, mesiolingual area, distobuccal area, distolingual area, mesial marginal ridge, and distal marginal ridge. Image measurement software (3Shape Unite) was employed to record the smallest preparation space on the molar occlusal surface, and a space of <1 mm was defined as insufficient preparation. The chi-square test was utilized for the statistical analysis of data. RESULTS: Significant differences were observed in the areas with the smallest preparation space on the occlusal surface of molars across different quadrants (P<0.05). Notably, the area with the smallest preparation space on the occlusal surface of the maxillary first molar was most frequently found in the mesiobuccal area, whereas the smallest preparation spaces on other molars were predominantly located in the distolingual area. The insufficient preparation of the occlusal surface occurred in 41.0% (347 cases) of teeth. The distribution of prone areas corresponded with the distribution of the smallest spatial areas. CONCLUSIONS Although the fixed-depth groove method is employed, the preparation space on the molar surface unavoidably remains uneven under visual guidance. The identification of the areas prone to minimum preparation spaces can serve as a valuable guide for clinicians to prevent insufficient preparation.
Humans ; Molar/anatomy & histology* ; Crowns ; Zirconium

Objective:To study the protective effect of Pien-Tze-Huang on acetaminophen-induced liver injury,and to clarify the possible mechanism.Methods:The human normal hepatocytes(L02 cells)were divided into control group,APAP group(10 mmol·L-1 APAP),APAP+PZH group(10 mmol·L-1 APAP and 0.4 mg·mL-1 PZH),and PZH group(0.4 mg·mL-1 PZH).The survival rates of the cells in various groups were determined by MTT method,the morphology was observed by inverted microscope,and the apoptotic rates were detected by flow cytometry.The activities of superoxide dismutase(SOD)and lactate dehydrogenase(LDH)and the levels of malondialdehyde(MDA)in the cell supernant in various groups were detected by the kits,the reactive oxygen species(ROS)levels and the mitochondrial membrane potential(MMP)in the hepatocytes in various groups were detected by fluorescence probe.Western blotting method was used to examine the expression levels of apoptosis-related proteins,phosphatidylinositol 3 kinase(PI3K)/protein kinase B(AKT)signaling pathway proteins,nuclear factor-κB(NF-κB)signaling pathway proteins and inflammatory factors in the cells in various groups.Results:The results of MTT showed that compared with control group,the survival rate of the cells in APAP group was markedly decreased(P＜0.05);compared with APAP group,the survival rate of the cells in APAP+PZH group was significantly increased(P＜0.05).Compared with control group,the number of the L02 cells in APAP group showed a decreasing and loosely arranged trend;compared with APAP group,the number and arrangement of the L02 cells in APAP+PZH group were notably improved.The results of flow cytometry showed that compared with control group,the apoptotic rate of the L02 cells in APAP group was significantly increased(P＜0.05);compared with APAP group,the apoptotic rate of the cells in APAP+PZH group was significantly decreased(P＜0.05).Compared with control group,the activity of LDH and level of MDA in the cells in APAP group were significantly increased(P＜0.05),while the activity of SOD was significantly decreased(P＜0.05);compared with APAP group,the activity of LDH and level of MDA in the cells in APAP+PZH group were significantly decreased(P＜0.05),while the activity of SOD was significantly increased(P＜0.05).Compared with control group,the fluorescence intensity of ROS in the cells in APAP group was significantly increased;compared with APAP group,the fluorescence intensity of ROS in APAP+PZH group was significantly decreased.Compared with control group,the MMP of the L02 cells in APAP group was significantly decreased;compared with APAP group,the MMP of the L02 cells in APAP+PZH group was significantly increased.The results of Western blotting showed that compared with control group,the expression levels of caspase-9,B-cell lymphoma 2(Bcl-2)associated X protein(BAX),phosphorylated PI3K(p-PI3K),phosphorylated AKT(p-AKT),phosphorylated NF-κB inhibitor alpha(p-IKBα),p-P65,phosphorylated inhibitor of kappaB kinaseβ(p-IKKβ),interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)proteins in the cells in APAP group were significantly increased(P＜0.05),while the level of Bcl-2 proteins in the cells in APAP group was significantly decreased(P＜0.05);compared with APAP group,the levels of caspase-3,BAX,p-PI3K,p-AKT,p-IKBα,p-P65,p-IKKβ,IL-1β,IL-6,and TNF-α proteins in the cells in APAP+PZH group were significantly decreased,while the level of Bcl-2 protein was significantly increased(P＜0.05).Conclusion:PZH may reduce the oxidative stress and inflammatory response in the cells by regulating the PI3K/AKT and NF-κB signaling pathway,therefore attenuate the L02 cell injury induced by APAP.

Objective To investigate the effects of downregulating microRNA(miR)-550a-5p on the proliferation,migration and invasion of non-small cell lung cancer(NSCLC)and to explore its mo-lecular mechanisms.Methods The miR-550a-5p inhibitor and its negative control(inhibitor NC),as well as small interfering RNA targeting scavenger receptor class A member 3(si-SCARA3)and its negative control(si-NC)were individually or co-transfected into A549 cells.These cells were desig-nated as inhibitor NC group,miR-550a-5p inhibitor group,miR-550a-5p inhibitor+si-NC(co-trans-fection)group and miR-550a-5p inhibitor+si-SCARA3(co-transfection)group.Cell proliferation was assessed by CCK-8 assay;colony formation ability was evaluated using a plate cloning method;cell migration and invasion were detected by scratch assay and Transwell chamber assay,respectively;levels of matrix metalloproteinase(MMP)-2 and MMP-9 in the supernatant of A549 cells were measured by enzyme-linked immunosorbent assay(ELISA);apoptosis levels were determined by Annexin V/PI double staining.Results Compared with adjacent normal tissues,the expression level of miR-550a-5p was increased,while SCARA3 mRNA expression was decreased in NSCLC tissues(P＜0.05).Dual-luciferase reporter assays confirmed that miR-550a-5p directly targeted SCARA3.Compared with the inhibitor NC group,cell proliferation rate,colony formation number,migration and invasion ability of the miR-550a-5p inhibitor group decreased,apoptosis rate increased(P＜0.05).Compared with the inhibitor NC group,the expression levels of MMP-2 and MMP-9 in cell supernatant of the miR-550a-5p inhibitor group were decreased,and the expression levels of SCARA3 protein were increased(P＜0.05).Compared with the miR-550a-5p inhibitor group or the miR-550a-5p inhibitor+si-NC group,cell proliferation rate,clonal colony formation number,migration and invasion ability of the miR-550a-5p inhibitor+si-SCARA3 group were increased,and apoptosis rate was decreased(P＜0.05).Compared with the miR-550a-5p inhibitor group or the miR-550a-5p inhibitor+si-NC group,the miR-550a-5p inhibitor+si-SCARA3 group showed in-creased expression levels of MMP-2 and MMP-9 in the cell supernatant and decreased SCARA3 pro-tein expression levels(P＜0.05).Conclusion Downregulation of miR-550a-5p inhibits the prolif-eration,migration and invasion of NSCLC cells,and promotes apoptosis,possibly through upregulat-ing SC AR A3 expression.

Objective To explore the relationship between serum miRNA-21 and miR-27b levels and prognosis of patients with renal clear cell carcinoma.Methods A total of 118 patients with renal clear cell carcinoma admitted to the Qinghai University Hospital from February 2019 to April 2021 were selected as the study subjects,and another 118 healthy patients in the same period as the control group.Real time fluorescence quantitative polymerase chain reaction(PCR)was used to detect the expression of miR-21 and miR-27b in the serum of all subjects.The relative expression levels of serum miR-21 and miR-27b between the patients with renal clear cell carcinoma and healthy control patients were compared.The expression and correlation of serum miR-21 and miR-27b in the patients with renal clear cell carcinoma of different pathological stages and Fuhrman grading were analyzed.The relationship between the expression of serum miR-21 and miR-27b and the survival and prognosis of the patients was explored as well.Results The expression levels of serum miR-21 and miR-27b in the patients with renal clear cell carcinoma were higher than those in the healthy control group(P＜0.05).The serum miR-21 expression level in stage Ⅲ patients was higher than in stageⅠ(P＜0.05),while the serum miR-21 expression level in the stage Ⅳ patients was higher than that in stagesⅠ,Ⅱ,and Ⅲ(P＜0.05).The expression level of miR-27b in the serum of patients gradually increased across the four stages,with a significant difference(P＜0.05).The pathological staging was positively correlated with the expression of miR-21 and miR-27b(P＜0.001).The expression levels of miR-21 and miR-27b in serum of patients gradually increased across grades Ⅰ,Ⅱ and Ⅲ by Fuhrman grading,with significant difference(P＜0.05).Fuhrman grading was positively correlated with the serum miR-21 and miR-27b expression(P＜0.001).There was a statistically significant difference in the survival curve between the miR-21 high expression group and the low expression group(P＜0.05).There was a statistically significant difference in the survival curve between the high expression group and the low expression group of miR-27b(P＜0.05).Conclusion The expression levels of serum miR-21 and miR-27b in patients with renal clear cell carcinoma is indicative of the progression and prognosis of the patient's condition.

With rapid development of information technology,blended teaching model has gradually become mainstream teaching model in most colleges and universities.How to evaluate students'learning effect and analyze factors that affect students'per-formance is a key research direction of this teaching model.Taking blended teaching of Medical Immunology course in Xinxiang Medical University as an example,this paper introduces implementation process and evaluation system of blended teaching,analyzes main factors affecting students'performance and learning effect,and focuses on discussing influence of formative evaluation and"flipped classroom"on students'academic performance.

Objective:To evaluate the effectiveness of the artificial intelligence-assisted diagnosis and treatment system in distinguishing benign and malignant lung nodules.Methods:Clinical data of 1 650 patients with pulmonary nodules admitted to the Tianjin Chest Hospital Affiliated to Tianjin University from January 2015 to October 2022 were retrospectively analyzed, which included a total of 1 650 nodules. The malignant probability was compared to and analyzedpredict the benign and malignant nodules.Results:A total of 1 650 nodules were included, which were divided into a malignant tumor group(n=1 391, 84.3%) and a benign lesion group(n=259, 15.7%). Between the malignant tumor group and the benign lesion group, the malignant probability was significantly different, and the malignant probability could better distinguish malignant nodules and benign lesions[(78.85±22.91)% vs.(54.91±28.68)%, P<0.001]. The area under the curve( AUC) was 0.768. The critical value of malignant probability for diagnosis of lung cancer was 81.3% with a sensitivity of 0.620 and specificity of 0.815. Stratified analysis results showed that the accuracy of the AI intelligent system for diagnosing pulmonary nodules with the sizes of 0-1 cm, 1-2 cm and 2-3 cm was also increased, and the areas under ROC curve were 0.717, 0.769 and 0.804, respectively. Conclusion:The malignant probability of lung nodules calculated by the artificial intelligence-assisted diagnosis and treatment system can be used in the assessment of the preoperative benign and malignant lung nodules.

Objective:To analyze the clinical characteristics of elderly acute pulmonary thromboembolism(APE)patients complicated with preexisting atrial fibrillation(AF)and the impact of preexisting AF on in-hospital adverse outcomes in elderly patients with APE.Methods:A retrospective analysis was performed on elderly APE patients with preexisting AF hospitalized in Beijing Anzhen Hospital, Capital Medical University between January 1, 2008 and December 31, 2021.We compared the comorbidities, symptoms, signs, laboratory test results and echocardiographic features, simplified pulmonary embolism severity index(sPESI)scores and adverse in-hospital outcomes between the preexisting AF group and the non-AF group.Logistic regression was used to analyze the risk factors of in-hospital adverse outcomes in elderly patients with APE.Results:A total of 240 patients diagnosed with APE were enrolled.There were 120 patients in the AF group and 120 patients in the non-AF group.For patients in the AF group and the non-AF group, the proportions with chronic heart failure were 38.3%(46/120)and 15.8%(19/120), the proportions with lower extremity deep vein thrombosis(DVT)were 36.7%(44/120)and 65.8%(79/120), the left ventricular ejection fractions(LVEF)were(59±10)% and(62±7)%, and hospital stays were(15±7)and(11±4)days, respectively, and the differences were statistically significant( χ2=15.381, 20.429, t=2.527, -4.710, all P<0.05). The incidences of in-hospital adverse outcomes in the AF group and the non-AF group were 4.2%(5/120)and 3.3%(4/120), respectively, with no significant difference( χ2=0.000, P=1.000). The overall incidence of in-hospital adverse outcomes was 3.8%(9/240). Multivariate Logistic regression analysis showed that elevated lactic acid was an independent risk factor for in-hospital adverse outcomes( OR=2.753, 95% CI: 1.367-5.542, P=0.005). However, AF( OR=2.880, 95% CI: 0.587-14.141, P=0.192)and sPESI score( OR=2.056, 95% CI: 0.904-4.673, P=0.086)were not associated with in-hospital adverse outcomes. Conclusions:Elderly APE patients with preexisting AF have a relatively low incidence of DVT, but a higher proportion have concurrent chronic heart failure and need a longer hospital stay.Elevated lactic acid is an independent risk factor for in-hospital adverse outcomes of elderly APE patients with preexisting AF.However, preexisting AF has no predictive value for in-hospital adverse outcomes in elderly patients with APE.

CRISPR-Associated Protein 9 ; Gene Editing ; CRISPR-Cas Systems/genetics*

To investigate the clinical role of T-cell transcription factor (TBX21) and adenylate cyclase 9 antibody (ADCY9) gene polymorphisms in the development of childhood asthma. METHODS: Two hundred Han Chinese wheezing children aged 5 years and younger in Henan region from July 2016 to January 2017 were selected as the study group, and another 100 Han Chinese healthy children aged 5 years and younger in the same period were selected as the control group. Oral mucosal exfoliated cells were collected from both groups, and the genotypes of TBX21 gene rs2240017 polymorphic locus and ADCY9 gene rs2230739 polymorphic locus were detected by real-time fluorescence quantitative polymerase chain reaction (PCR) technique, and the risk level of asthma was assessed based on the test results. The children in the low-risk and high-risk groups were compared in terms of serum immunoglobulin E (IgE) levels, API positivity rate and allergic disease incidence, and the correlation between the risk level of asthma-related genetic polymorphisms and serum IgE levels, API and allergic disease incidence was analyzed. All children were followed up until 6 years of age to confirm the diagnosis of asthma, and the incidence of asthma was compared between the low-risk and high-risk groups. Children with asthma were treated with inhaled glucocorticoids and leukotriene receptor antagonists for 3 months, and the control of asthma and the impairment of lung function were compared between the low-risk and high-risk groups. RESULTS: The genotype detection results of rs2240017 polymorphic locus of TBX21 gene and rs2230739 polymorphic locus of ADCY9 gene in the study group compared with those in the control group were statistically significant (P<0.001). The percentages of CC, CT, and TT genotypes of rs2240017 polymorphic locus of TBX21 gene were 19.50%, 56.00%, and 24.50%, respectively, and the percentages of CC, CG, and GG genotypes of rs2230739 polymorphic locus of ADCY9 gene were 86.00%, 10.00%, and 4.00%, respectively, in 200 children with wheezing; serum IgE level, API positivity rate and allergic disease incidence were higher in the high-risk group than in the low-risk group (P< 0.001, <0.001, 0.021, respectively). The degree of risk of asthma-related gene polymorphisms in children with wheezing was positively correlated with serum IgE levels, API positivity, and the incidence of allergic diseases (P<0.001); the incidence of asthma (81.48%) and impaired lung function (74.07%) were higher in the high-risk group than in the low-risk group (4.90%, 3.50%) (P<0.001). There was no statistically significant difference between the asthma control rate of children with asthma in the high-risk group (79.55%) compared with the asthma control rate of children with asthma in the low-risk group (100.00%) (P=0.433). CONCLUSION: Gene polymorphisms at rs2240017 locus of TBX21 gene and rs2230739 locus of ADCY9 gene are closely associated with asthma development and impaired lung function in children with wheezing.