Analysis of intraoperative frozen section diagnosis of 1 263 pulmonary nodules / 中国胸心血管外科临床杂志

@#Objective    To explore the key points and difficulties of intraoperative frozen section diagnosis of pulmonary diseases. Methods    The intraoperative frozen section and postoperative paraffin section results of pulmonary nodule patients in Beijing Chaoyang Hospital, Capital Medical University from January 2021 to January 2022 were collected. The main causes of misdiagnosis in frozen section diagnosis were analyzed, and the main points of diagnosis and differential diagnosis were summarized. Results    According to the inclusion criteria, a total of 1 263 frozen section diagnosis results of 1 178 patients were included in the study, including 475 males and 703 females, with an average age of 58.7 (23-86) years. In 1 263 frozen section diagnosis results, the correct diagnosis rate was 95.65%, and the misdiagnosis rate was 4.35%. There were 55 misdiagnoses, including 18 (3.44%) invasive adenocarcinoma, 17 (5.82%) adenocarcinoma in situ, 7 (35.00%) mucinous adenocarcinoma, 4 (2.09%) minimally invasive adenocarcinoma, 3 (100.00%) IgG4 related diseases, 2 (66.67%) mucinous adenocarcinoma in situ, 1 (16.67%) atypical adenomatous hyperplasia, 1 (14.29%) sclerosing pulmonary cell tumor, 1 (33.33%) bronchiolar adenoma, and 1 (100.00%) papillary adenoma. Conclusion    Intraoperative frozen section diagnosis still has its limitations. Clinicians need to make a comprehensive judgment based on imaging examination and clinical experience.

Exploring the mechanism of Xihuang Pill's anti-hyperplasia of mammary glands effect based on UPLC-Q-orbitrap-MS technology / 药学学报

Based on UPLC-Q-orbitrap-MS and biological network analysis tools, the mechanism of Xihuang Pill in improving hyperplasia of mammary glands was systematically analyzed. The rat model of hyperplasia of mammary glands was established by intramuscular injection of estradiol benzoate and progesterone. LC-MS tissue metabolomics was used to explore the key metabolites and metabolic pathways of Xihuang Pill in improving hyperplasia of mammary glands in rat. The network analysis of the key metabolites regulated by Xihuang Pill was carried out by integrating biological network analysis tools, focusing on the key metabolic pathways, and exploring the potential targets of Xihuang Pill to improve hyperplasia of mammary glands. Compared with the control group, there were significant differences in the content of 49 differential metabolites in the tissues of the model group (P < 0.05). Xihuang Pills could significantly call back 17 metabolites such as L-alanine, threonine, indole-3-carboxylic aldehyde, lysine, arginine, alanylleucine, glycyltyrosine, γ-glutamyl leucine, vitamin B3, serine leucine, threonine leucine, isoleucine glutamic acid, γ-glutamyl tyrosine, decanoyl-L-carnitine, uric acid, leucylleucine, S-adenosyl-methionine. Further network analysis and literature research on the key metabolites regulated by Xihuang Pills showed that the AGE-RAGE signaling pathway may be one of the important pathways for Xihuang Pills to improve hyperplasia of mammary glands. STAT3, MAPK1, EGFR, CASP3, CASP8, PRKCA and JUN in the AGE-RAGE signaling pathway may be potential targets for Xihuang Pills to improve hyperplasia of mammary glands. The animal experiment operations involved in this paper follow the provisions of the Animal Ethics Committee of Gansu University of Traditional Chinese Medicine and pass the ethical review of animal experiments (approval number: 2022-705).

Study on quality control method of the roots and rhizoma of Toricellia angulata / 中国药房

OBJECTIVE To establish the quality control method for the roots and rhizoma of Toricellia angulata. METHODS The properties of the roots and rhizoma of T. angulata were observed and microscopic identification was conducted. The moisture， total ash， acid-insoluble ash and ethanol-soluble extract were examined according to the method stated in the 2020 edition of Chinese Pharmacopoeia （part Ⅳ）. HPLC fingerprints of 11 batches of the roots and rhizoma of T. angulata were established， common peaks were identified and the similarity was evaluated by using the Similarity Evaluation System of Chromatographic Fingerprint of TCM （2012 edition）. The contents of coniferin， syringin， chlorogenic acid， （+）-syringaresinol-O-β-D-glucopyranoside and syringaresinol were determined by HPLC. RESULTS The properties and microscopic identification of the roots and rhizoma of T. angulata were obvious. The average contents of moisture， total ash， acid-insoluble ash and ethanol-soluble extract were 7.54%， 2.18%， 0.15% and 7.81%， respectively. There were 16 common peaks marked in the HPLC fingerprints of 11 batches of the roots and rhizoma of T. angulata， with similarities of 0.856-0.960； five of them were identified， such as coniferin， syringin， chlorogenic acid， （+）-syringaresinol-O-β-D-glucopyranoside and syringaresinol. The contents of the above five components were 0.047 2-0.401 6， 0.836 8-8.697 9， 1.245 3-10.950 0， 0.139 0-0.437 8 and 0.016 4-0.635 3 mg/g， respectively. CONCLUSIONS The established method is stable and accurate， which can be used for the quality control of the roots and rhizoma of T. angulata. It is preliminarily proposed that the moisture in the roots and rhizoma of T. angulata is not more than 11.0%， the total ash is not more than 4.0%， the ethanol-soluble extract is not less than 5.0%， the contents of coniferin， syringin， chlorogenic acid， （+）-syringaresinol-O-β-D- glucopyranoside and syringaresinol are not less than 0.04，0.83， 1.24， 0.13， 0.01 mg/g， respectively.

Association between clinical phenotypes of hypertrophic cardiomyopathy and Ca2+ gene variation gene variation / 中华心血管病杂志

Objective: To observe the association between clinical phenotypes of hypertrophic cardiomyopathy (HCM) patients and a rare calcium channel and regulatory gene variation (Ca2+ gene variation) and to compare clinical phenotypes of HCM patients with Ca2+ gene variation, a single sarcomere gene variation and without gene variation and to explore the influence of rare Ca2+ gene variation on the clinical phenotypes of HCM. Methods: Eight hundred forty-two non-related adult HCM patients diagnosed for the first time in Xijing Hospital from 2013 to 2019 were enrolled in this study. All patients underwent exon analyses of 96 hereditary cardiac disease-related genes. Patients with diabetes mellitus, coronary artery disease, post alcohol septal ablation or septal myectomy, and patients who carried sarcomere gene variation of uncertain significance or carried>1 sarcomere gene variation or carried>1 Ca2+ gene variation, with HCM pseudophenotype or carrier of ion channel gene variations other than Ca2+ based on the genetic test results were excluded. Patients were divided into gene negative group (no sarcomere or Ca2+ gene variants), sarcomere gene variation group (only 1 sarcomere gene variant) and Ca2+ gene variant group (only 1 Ca2+ gene variant). Baseline data, echocardiography and electrocardiogram data were collected for analysis. Results: A total of 346 patients were enrolled, including 170 patients without gene variation (gene negative group), 154 patients with a single sarcomere gene variation (sarcomere gene variation group) and 22 patients with a single rare Ca2+ gene variation (Ca2+ gene variation group). Compared with gene negative group, patients in Ca2+ gene variation group had higher blood pressure and higher percentage of family history of HCM and sudden cardiac death (P<0.05); echocardiographic results showed that patients in Ca2+ gene variation group had thicker ventricular septum ((23.5±5.8) mm vs. (22.3±5.7) mm, P<0.05); electrocardiographic results showed that patients in Ca2+ gene variation group had prolonged QT interval ((416.6±23.1) ms vs. (400.6±47.2) ms, P<0.05) and higher RV5+SV1 ((4.51±2.26) mv vs. (3.50±1.65) mv, P<0.05). Compared with sarcomere gene variation group, patients in Ca2+ gene variation group had later onset age and higher blood pressure (P<0.05); echocardiographic results showed that there was no significant difference in ventricular septal thickness between two groups; patients in Ca2+ gene variation group had lower percentage of left ventricular outflow tract pressure gradient>30 mmHg (1 mmHg=0.133 kPa, 22.8% vs. 48.1%, P<0.05) and the lower early diastolic peak velocity of the mitral valve inflow/early diastolic peak velocity of the mitral valve annulus (E/e') ratio ((13.0±2.5) vs. (15.9±4.2), P<0.05); patients in Ca2+ gene variation group had prolonged QT interval ((416.6±23.1) ms vs. (399.0±43.0) ms, P<0.05) and lower percentage of ST segment depression (9.1% vs. 40.3%, P<0.05). Conclusion: Compared with gene negative group, the clinical phenotype of HCM is more severe in patients with rare Ca2+ gene variation; compared with patients with sarcomere gene variation, the clinical phenotype of HCM is milder in patients with rare Ca2+ gene variation.

Construction of core competence evaluation index system for in-service nurses in the imaging department / 中华医学教育探索杂志

Objective:To construct an evaluation index system for core competence of nurses in the imaging department.Methods:The literature research and the opinions of the research team were integrated to develop indicators for core competence of imaging nurses at all levels. Delphi method was used to conduct 2 rounds of consultation with 20 experts, and analytic hierarchy process (AHP) was used to determine the index weights at all levels. The indicators were converted into a self-assessment scale, and 31 imaging in-service nurses in a third-level hospital were selected for questionnaire survey. SPSS 21.0 software was used to analyze the consultation results.Results:Through 2 rounds of consultation, the evaluation index system for core competence of nurses in the imaging department was formed, including 5 first-level indicators, 18 second-level indicators, and 78 third-level indicators. The effective response rates of the two rounds of expert consultation were 100%, and the authority coefficients were 0.933 and 0.923. In the second round, the expert opinion coordination coefficients of the first, second and third grade indexes were 0.46, 0.53 and 0.47, respectively ( P < 0.01). The core competence scores of the 31 respondents ranged from 3.08 to 4.45 in each dimension. Conclusion:After 2 rounds of expert consultation, the expert opinions tended to be consistent and the degree of coordination was high. The constructed indicators can be used for the core competence training and teaching evaluation of in-service nurses in the imaging department.

Research on the quality certification standard of medical MOOC in universities / 中华医学教育探索杂志

The medical massive open online course (MOOC) quality certification standard is the reference for the implementation of medical MOOC quality monitoring. This thesis utilizes data mining, quantitative analysis, qualitative analysis and other methods, taking the comment text of the Chinese medicine courses on the Ai Course (MOOC of China Universities) platform as the data source out of which to extract certain potential indicators for the medical MOOC quality certification standard. Based on the analysis of the existing research results on MOOC quality evaluation domestic and abroad as well as the relevant policies of the Ministry of Education, a medical MOOC quality accreditation standard including 6 first-level indicators are formed, including course content, teaching design and so on. Expert scores and AHP method were used to calculate the weight coefficient of the quality certification standard. Finally, this paper explains the procedure of medical MOOC quality certification in college. This certification system can be used to conduct self-assessment and quality monitoring of medical MOOC in colleges and universities.

Dose-adjusted concentrations of Posaconazole oral suspension in hematopoietic stem cell transplantation patients and analysis of the influential factors / 中国药房

OBJECTIVE To analyze the dose-adjusted concentrations of Posaconazole oral suspension in patients undergoing hematopoietic stem cell transplantation （HSCT） and their influential factors. METHODS Data were collected from hospitalized HSCT patients admitted to the First Affiliated Hospital of Shandong First Medical University （Shandong Provincial Qianfoshan Hospital） from January 2021 to April whtwhm@yeah.net 2023 who took Posaconazole oral suspension for the prevention of invasive fungal disease （IFD） and received blood concentration of posaconazole. The rate of concentration attainment and clinical failure rate of posaconazole for the prevention of IFD were evaluated， and one-way and multiple linear regression analyses were performed for the influential factors of dose-adjusted concentrations （C0/D） of posaconazole. RESULTS A total of 44 patients were enrolled； the mean C0 of posaconazole in patients was （0.99±0.94） µg/mL， and 20 patients had a C0≥0.7 μg/mL， with a concentration attainment rate of 45.45% for the prevention of IFD； 13 cases were clinical failures， with a clinical failure rate of 29.55%. Of 24 patients who did not achieve C0/D of posaconazole for IFD prophylaxis， one patient was a clinical failure despite timely dose adjustment of posaconazole in seven patients； seven of the thirteen patients who did not undergo dose adjustment were clinical failures； and the remaining four patients were switched to other antifungal agents. The results of univariate analysis showed that gender， body mass index （BMI）， renal function， combined use of sodium phenytoin， omeprazole and metoclopramide had a significant effect on the C0/D of posaconazole （P＜0.05）； the results of multivariate linear regression analysis showed that gender， BMI and combined use of sodium phenytoin were the independent factors affecting the C0/D of posaconazole （P＜0.05）. CONCLUSIONS Significant individual differences are reflected in the blood concentration of Posaconazole oral suspension； gender， BMI and combined use of sodium phenytoin are independent factors affecting the C0/D of posaconazole.

Effects of KCNQ1OT1 Gene Knockout Combined with Bruceine D on Proliferation, Migration, and Invasion of Breast Cancer MDA-MB-231 Cells / 肿瘤防治研究

Objective To explore the effect of KCNQ1OT1 gene knockout combined with bruceine D on the proliferation, migration, and invasion of breast cancer MDA-MB-231 cells. Methods Cell Counting Kit-8, wound healing, and Transwell invasion assay were used to detect the effects of bruceine D and siKCNQ1OT1 on the viability, migration, and invasion of MDA-MB-231 cells. Effect of bruceine D and siKCNQ1OT1 on the expression of KCNQ1OT1 in MDA-MB-231 cells was detected by qRT-PCR. Western blot was used to detect the effect of bruceine D and siKCNQ1OT1 on the expression of EMT-related proteins and CDC42, p-MKK7, MKK7 proteins in MDA-MB-231 cells. Results Bruceine D and siKCNQ1OT1 could significantly inhibit the viability, migration, and invasion of MDA-MB-231 cells, and the inhibitory effect was enhanced when they were combined (all P < 0.05); bruceine D downregulated the expression of KCNQ1OT1 in MDA-MB-231 cells (all P < 0.05); bruceine D combined with siKCNQ1OT1 significantly decreased CDC42, p-MKK7, N-cadherin, and Vimentin expression in MDA-MB-231 cells and increased the expression of E-cadherin (all P < 0.05). Conclusion Bruceine D combined with siKCNQ1OT1 significantly inhibit the proliferation, migration, invasion, and EMT of human breast cancer MDA-MB-231 cells, and its molecular mechanism may be related to the blocking of CDC42/MKK7 signaling pathway.

Feasibility study on rapid identification of methicillin-resistant Staphylococcus aureus by different algorithms of MALDI-TOF mass spectrometry / 中华检验医学杂志

Objective:To explore the feasibility of rapid identification of methicillin-resistant Staphylococcus aureus using different algorithms of the matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometer. Methods:Totally 314 clinical isolates of Staphylococcus aureus were selected from the bacterial bank at Beijing Tongren Hospital from January 2017 to June 2019. The samples were identified by MALDI-TOF MS, and screened by cefoxitin disk method (inhibition ring diameter ￡21 mm) and PCR mecA gene. The strains were divided into a methicillin-resistant Staphylococcus aureus (MRSA) group (130 strains) and a methicillin-susceptible Staphylococcus aureus (MSSA) group (184 strains). Then, after collecting the spectrograms of these samples using formic acid extraction, the MRSA group and MSSA group were divided into three subgroups each, namely MRSA-1 (43 strains), MRSA-2 (42 strains), MRSA-3 (45 strains) and MSSA-1 (60 strains), MSSA-2 (61 strains) and MSSA-3 (63 strains). The groups were studied using genetic algorithm (GA), fast classification algorithm (QC) and supervised neural network algorithm (SNN) in the ClinProTools software on the Bruker MALDI-TOF mass spectrometer, and the convolutional neural network algorithm (CNN) in the Ex-SmartSpec software on the Zhongyuan Hui-Ji mass spectrometer. These studies were repeated for 3 rounds. The first round with MRSA-1 and MRSA-2, MSSA-1 and MSSA-2 being model groups, MRSA-3 and MSSA-3 being validation groups. The validation groups were rotated for each round. The areas under the receiver operating characteristic (ROC) curve expansions of the four algorithms were used to confirm each program′s performance. Then, 38 MRSA strains and 40 MSSA clinical strains were selected from the bacterial bank of the Laboratory of Beijing Tongren Hospital from July 2019 to December 2019, and were put through the formic acid extraction method to collect their spectra. These samples were tested independently with their convolutional neural network models. Results:After three rounds of modeling and verification, the areas under the ROC curves of the three Bruker ClinProTools programs were as follows: for genetic algorithm, the areas were 0.89, 0.74, and 0.64 respectively; for fast classification algorithm, the areas were 0.77, 0.95, and 0.94 respectively; and for supervised neural network algorithm, the areas were 0.90, 0.98, and 0.98 respectively. The areas under the ROC curves of the convolutional neural network algorithm with Zhongyuan Huiji mass spectrometer′s Ex-SmartSpec software were 0.95, 0.99, and 0.99 respectively. The independent test results of convolutional neural network algorithm showed that these results have an accuracy, specificity, sensitivity and AUC of 88.82% (810/912), 81.15% (779/960), 84.88% (1 589/1 872) and 0.92 respectively.Conclusions:The supervised neural network algorithm of Bruker′s ClinProTools and the convolutional neural network algorithm of Zhongyuan Hui-Ji mass spectrometer′s EX-Smartspec is clinically acceptable for rapid identification of MRSA performance indicators. Using convolutional neural network algorithm and MALDI-TOF mass spectrometry, MRSA strains can be identified quickly, providing timely advice for clinical medications.

Exercising to music combined with transcranial direct current stimulation improves the motor and cognitive functioning of persons with Parkinson′s disease / 中华物理医学与康复杂志

Objective:To observe any effect of combining music exercise with transcranial direct current stimulation (tDCS) on the motor control, balance and cognition of persons with Parkinson′s disease (PD).Methods:A total of 120 PD patients were randomly divided into a control group, a music exercise group, a tDCS group and a combined group, each of 30. All received routine rehabilitation training, while the music exercise, tDCS and combined groups were additionally provided with music exercise therapy, tDCS treatment or both, respectively. Version three of the unified Parkinson′s disease scale (UPDRSIII), a 10m reentry movement test, the Berg balance scale (BBS), the Activity Balance Confidence scale (ABC) and Montreal cognitive assessments were applied before and after 4 weeks of the treatments.Results:After the treatment, the average UPDRSIII score and 10m reentry movement time of the music exercise group were significantly lower than in the control group, while the average BBS and ABC scores were significantly higher than the control group′s averages. The tDCS group′s average MoCA scores on all of the items and its total score were significantly higher than those of the music exercise and control groups. The average UPDRSIII score and 10m reentry movement time of the combined group were the lowest after the treatment, and that group′s average BBS, ABC, MoCA and total scores were the highest, significantly better than the other three groups.Conclusion:Combining music exercise training with tDCS can effectively improve the motor functioning, balance and cognition of persons with PD.

Supplementing transcranial magnetic stimulation with intermittent resistance training improves the recovery of motor function in Parkinson′s disease / 中华物理医学与康复杂志

Objective:To explore any effect of supplementing low-frequency repeated transcranial magnetic stimulation (rTMS) with intermittent resistance training on muscle tone and the recovery of motor function among persons with Parkinson′s disease (PD).Methods:A total of 104 PD patients were randomly divided into an observation group and a control group, each of 52. Both groups were treated with conventional drugs and low-frequency rTMS, while the observation group was additionally provided with intermittent resistance training for 8 weeks. Before and after the intervention, the motor functioning of both groups was evaluated using unified Parkinson′s disease rating scale-III (UPDRS-III) and the Berg Balance Scale (BBS). Limb muscle tension was quantified using the modified Ashworth scale (MAS). The subjects′ psychological states were quantified using the exercise self-efficacy scale (ESE), and the modified Barthel index (MBI) was applied to evaluate their ability in the activities of daily living. A 3D motion processing system collected and analyzed data describing each subject′s gait kinematics.Results:After the intervention, significantly greater average improvement was observed in all of the outcome measures among the observation group compared with the control group.Conclusion:Combining intermittent resistance training with low-frequency rTMS can significantly reduce muscle tone and improve the motor functioning of PD patients. The combination is more effective than low-frequency rTMS alone.

Supplementing rehabilitation training with botulinum neurotoxin improves outcomes for Parkinson′s disease patients with striatal foot deformity / 中华物理医学与康复杂志

Objective:To observe any therapeutic effect of combining botulinum toxin type A (BTX-A) with rehabilitation training in treating Parkinson′s disease (PD) patients with striatal foot deformity (SFD).Methods:A total of 68 PD patients with SFD were randomly divided into a control group and a treatment group. Both groups were given routine medication with pramipexole and dopamine receptor agonists and received lower limb rehabilitation training, including passive activity training, strength training and walking training. The treatment group was additionally injected with BTX-A. Sciatic pain was quantified using a visual analogue scale. The Unified Parkinson′s Disease Rating Scale-lower limb motor lower limb motor function (UPDRS-LLM) scale, the Berg balance scale and the modified Barthel index were applied to test all of the participants before the experiment and on the 7th, 14th and 30th day of the treatment.Results:The average scores of the control group on all of measures at were significantly better than those of the control group at the same time points, and by the 14th and 30th day had improved significantly compared with those before treatment.Conclusion:Supplementing rehabilitation training with BTX-A can significantly improve foot deformity and relieve the muscle tension and spastic pain of PD patients with SFD, promoting the motor functioning of their lower limbs, their balance and their performance in the activities of daily living.

Associations between serum GDF15 and glycolipid metabolism disorder in metabolic associated fatty liver patients / 中华内科杂志

Objective:To investigate relationships between serum growth differentiation factor 15 (GDF15) and glycolipid metabolism in patients with metabolic associated fatty liver disease (MAFLD).Methods:The current investigation was a cross-sectional study. A total of 333 patients from the Fengxian District Central Hospital were recruited into the study after physical examination from February 2020 to February 2021. There were 107 patients with MAFLD and type 2 diabetes mellitus (T2DM), including 54 males and 53 females with a mean age of (57±11) years. There were 65 patients with simple MAFLD only, including 32 men and 33 women with a mean age of (49±5) years. There were 105 patients with T2DM only, including 53 men and 52 women, with a mean age of (56±10) years. A control group of 56 people without MAFLD or diabetes,28 male, 28 female, mean age (48±6) years, was also included in the study. Serum GDF15 was measured via enzyme-linked immunosorbent assays. IBM SPSS 26.0 was used for statistical analysis. Logistic regression was used to evaluate relationships between GDF15 and metabolic abnormalities in MAFLD patients.Results:GDF15 progressively increased in the control [385 (296, 484) ng/L], nonobese MAFLD [388 (319, 435) ng/L], obese MAFLD [426 (354, 527) ng/L], T2DM [664 (483, 900) ng/L], and MAFLD+T2DM groups [770 (560, 1 074) ng/L]( H=113.82, P=0.001). There was no significant difference in serum GDF15 between the simple MAFLD [406 (339, 524) ng/L] and control group ( U=1 505.50, P=0.132). GDF15 was significantly higher in the MAFLD+T2DM group than in the T2DM-only group ( U=4 573.50, P=0.019). In logistic regression analysis increased GDF15 was associated with increased risks of simple MAFLD [odds ratio ( OR)=2.202], T2DM ( OR=29.656), and MAFLD+T2DM( OR=58.197). In patients with MAFLD, serum GDF15 was higher in the FIB4 index>1.45 group [773 (534, 1 162) ng/L] than in the FIB4 index<1.45 group [527 (389, 787) ng/L] ( U=1 709.50, P<0.001). Increased GDF15 was associated with an increased risk of advanced liver fibrosis ( OR=2.388). Conclusion:In patients with simple MAFLD, GDF15 level was not significantly higher than in the control group. In the T2DM-only group and the MAFLD+T2DM group GDF15 was significantly higher than in the control group. Increased serum GDF15 was associated with increased risk and severity of MAFLD complicated with abnormal glucose and lipid metabolism. High GDF15 increased the risk of advanced fibrosis in MAFLD patients.

Casticin Attenuates Stemness in Cervical Cancer Stem-Like Cells by Regulating Activity and Expression of DNMT1 / 中国结合医学杂志

OBJECTIVE@#To explore whether casticin (CAS) suppresses stemness in cancer stem-like cells (CSLCs) obtained from human cervical cancer (CCSLCs) and the underlying mechanism.@*METHODS@#Spheres from HeLa and CaSki cells were used as CCSLCs. DNA methyltransferase 1 (DNMT1) activity and mRNA levels, self-renewal capability (Nanog and Sox2), and cancer stem cell markers (CD133 and CD44), were detected by a colorimetric DNMT activity/inhibition assay kit, quantitative real-time reverse transcription-polymerase chain reaction, sphere and colony formation assays, and immunoblot, respectively. Knockdown and overexpression of DNMT1 by transfection with shRNA and cDNA, respectively, were performed to explore the mechanism for action of CAS (0, 10, 30, and 100 nmol/L).@*RESULTS@#DNMT1 activity was increased in CCSLCs compared with HeLa and CaSki cells (P<0.05). In addition, HeLa-derived CCSLCs transfected with DNMT1 shRNA showed reduced sphere and colony formation abilities, and lower CD133, CD44, Nanog and Sox2 protein expressions (P<0.05). Conversely, overexpression of DNMT1 in HeLa cells exhibited the oppositive effects. Furthermore, CAS significantly reduced DNMT1 activity and transcription levels as well as stemness in HeLa-derived CCSLCs (P<0.05). Interestingly, DNMT1 knockdown enhanced the inhibitory effect of CAS on stemness. As expected, DNMT1 overexpression reversed the inhibitory effect of CAS on stemness in HeLa cells.@*CONCLUSION@#CAS effectively inhibits stemness in CCSLCs through suppression of DNMT1 activation, suggesting that CAS acts as a promising preventive and therapeutic candidate in cervical cancer.

Comparison of three methods for establishing rat peri-implantitis model / 北京大学学报(医学版)

OBJECTIVE@#To compare the efficiency and effect of establishing rat peri-implantitis model by traditional cotton thread ligation and local injection of Porphyromonas gingivalis lipopolysaccharide (LPS) around the implant, as well as the combination of the two methods.@*METHODS@#Left side maxillary first molars of 39 male SD rats were extracted, and titanium implants were implanted after four weeks of healing. After 4 weeks of implant osseointegration, 39 rats were randomly divided into 4 groups. Cotton thread ligation (n=12), local injection of LPS around the implant (n=12), and the two methods combined (n=12) were used to induce peri-implantitis, the rest 3 rats were untreated as control group. All procedures were conducted under 5% isoflurane inhalation anesthesia. The rats were sacrificed 2 weeks and 4 weeks after induction through carbon dioxide asphyxiation method. The maxilla of the rats in the test groups were collected and marginal bone loss was observed by micro-CT. The gingival tissues around the implants were collected for further real time quantitative PCR (RT-qPCR) analysis, specifically the expression of tumor necrosis factor-alpha (TNF-α) as well as interleukin-1β (IL-1β). The probing depth (PD), bleeding on probing (BOP) and gingival index (GI) of each rat in the experimental group were recorded before induction of inflammation and before death.@*RESULTS@#After 4 weeks of implantation, the osseointegration of implants were confirmed. All the three test groups showed red and swollen gums, obvious marginal bone loss around implants. After 2 weeks and 4 weeks of inflammation induction, PD, GI and BOP of the three test groups increased compared with those before induction, but only BOP was statistically significant among the three test groups (P < 0.05). At the end of 2 weeks of inflammation induction, marginal bone loss was observed at each site in the cotton thread ligation group and the combined group. At each site, the bone resorption in the combined group was greater than that in the cotton thread ligation group, but the difference was not statistically significant (P > 0.05), bone resorption was observed at some sites of some implants in LPS local injection group. At the end of 4 weeks of inflammation induction, marginal bone loss was observed at all sites in each group. The marginal bone loss in the cotton thread ligation group and the combined group was greater than that in the LPS local injection group, and the difference was statistically significant (P < 0.05). At the end of 2 weeks and 4 weeks of induction, the expression of TNF-α and IL-1β in the test groups were higher than those in the control group (P < 0.05).@*CONCLUSION@#Compared with local injection of LPS around the implant, cotton thread ligature and the two methods combined can induce peri-implantitis in rats better and faster.

Correlation between pelvic relapses of rectal cancer after radical and R0 resection: A regression model-based analysis / 中华胃肠外科杂志

Objective: To propose a new staging system for presacral recurrence of rectal cancer and explore the factors influencing radical resection of such recurrences based on this staging system. Methods: In this retrospective observational study, clinical data of 51 patients with presacral recurrence of rectal cancer who had undergone surgical treatment in the Department of Gastrointestinal Surgery, Peking University People's Hospital between January 2008 and September 2022 were collected. Inclusion criteria were as follows: (1) primary rectal cancer without distant metastasis that had been radically resected; (2) pre-sacral recurrence of rectal cancer confirmed by multi-disciplinary team assessment based on CT, MRI, positron emission tomography, physical examination, surgical exploration, and pathological examination of biopsy tissue in some cases; and (3) complete inpatient, outpatient and follow-up data. The patients were allocated to radical resection and non-radical resection groups according to postoperative pathological findings. The study included: (1) classification of pre-sacral recurrence of rectal cancer according to its anatomical characteristics as follows: Type I: no involvement of the sacrum; Type II: involvement of the low sacrum, but no other sites; Type III: involvement of the high sacrum, but no other sites; and Type IV: involvement of the sacrum and other sites. (2) Assessment of postoperative presacral recurrence, overall survival from surgery to recurrence, and duration of disease-free survival. (3) Analysis of factors affecting radical resection of pre-sacral recurrence of rectal cancer. Non-normally distributed measures are expressed as median (range). The Mann-Whitney U test was used for comparison between groups. Results: The median follow-up was 25 (2-96) months with a 100% follow-up rate. The rate of metachronic distant metastasis was significantly lower in the radical resection than in the non-radical resection group (24.1% [7/29] vs. 54.5% [12/22], χ2=8.333, P=0.026). Postoperative disease-free survival was longer in the radical resection group (32.7 months [3.0-63.0] vs. 16.1 [1.0-41.0], Z=8.907, P=0.005). Overall survival was longer in the radical resection group (39.2 [3.0-66.0] months vs. 28.1 [1.0-52.0] months, Z=1.042, P=0.354). According to univariate analysis, age, sex, distance between the tumor and anal verge, primary tumor pT stage, and primary tumor grading were not associated with achieving R0 resection of presacral recurrences of rectal cancer (all P>0.05), whereas primary tumor pN stage, anatomic staging of presacral recurrence, and procedure for managing presacral recurrence were associated with rate of R0 resection (all P<0.05). According to multifactorial analysis, the pathological stage of the primary tumor pN1-2 (OR=3.506, 95% CI: 1.089-11.291, P=0.035), type of procedure (transabdominal resection: OR=29.250, 95% CI: 2.789 - 306.811, P=0.005; combined abdominal perineal resection: OR=26.000, 95% CI: 2.219-304.702, P=0.009), and anatomical stage of presacral recurrence (Type III: OR=16.000, 95% CI: 1.542 - 166.305, P = 0.020; type IV: OR= 36.667, 95% CI: 3.261 - 412.258, P = 0.004) were all independent risk factors for achieving radical resection of anterior sacral recurrence after rectal cancer surgery. Conclusion: Stage of presacral recurrences of rectal cancer is an independent predictor of achieving R0 resection. It is possible to predict whether radical resection can be achieved on the basis of the patient's medical history.

Activation of renal outer medullary potassium channel in the renal distal convoluted tubule by high potassium diet / 生理学报

Renal outer medullary potassium (ROMK) channel is an important K+ excretion channel in the body, and K+ secreted by the ROMK channels is most or all source of urinary potassium. Previous studies focused on the ROMK channels of thick ascending limb (TAL) and collecting duct (CD), while there were few studies on the involvement of ROMK channels of the late distal convoluted tubule (DCT2) in K+ excretion. The purpose of the present study was mainly to record the ROMK channels current in renal DCT2 and observe the effect of high potassium diet on the ROMK channels by using single channel and whole-cell patch-clamp techniques. The results showed that a small conductance channel current with a conductance of 39 pS could be recorded in the apical membrane of renal DCT2, and it could be blocked by Tertiapin-Q (TPNQ), a ROMK channel inhibitor. The high potassium diet significantly increased the probability of ROMK channel current occurrence in the apical membrane of renal DCT2, and enhanced the activity of ROMK channel, compared to normal potassium diet (P < 0.01). Western blot results also demonstrated that the high potassium diet significantly up-regulated the protein expression levels of ROMK channels and epithelial sodium channel (ENaC), and down-regulated the protein expression level of Na+-Cl- cotransporter (NCC). Moreover, the high potassium diet significantly increased urinary potassium excretion. These results suggest that the high potassium diet may activate the ROMK channels in the apical membrane of renal DCT2 and increase the urinary potassium excretion by up-regulating the expression of renal ROMK channels.

Causal relationship between branched-chain amino acids and peripheral atherosclerosis： a Mendelian randomization study / 上海预防医学

ObjectiveWe conducted a two-sample Mendelian randomization （MR） study to assess the causal relationship between circulating isoleucine， leucine and valine levels and the risk of peripheral atherosclerosis. MethodsBased on the large-scale genome-wide association study （GWAS） database， single nucleotide polymorphisms （SNPs） closely related to the circulating levels of isoleucine， leucine and valine were identified as instrumental variables （IVs）. Two-sample MR analysis applying the inverse variance weighted （IVW） method and the weighted median estimator （WME） method were performed to estimate the causal relationship between the risk of peripheral atherosclerosis and the exposure with more than three SNPs that were available as IVs. The pleiotropy was evaluated by using the MR-Egger regression and MR-PRESSO method， and the leave-one-out method was used in sensitivity analysis. ResultsFour， one and one SNPs were identified as IVs for circulating isoleucine， leucine and valine levels， respectively. For isoleucine， the IVW model demonstrated there was no evidence of heterogeneity among the IVs （P=0.715）， and there was a significant causal relationship between the increase of circulating isoleucine level and a higher risk of peripheral atherosclerosis risk. Per every 1 elevated standard deviation （SD） of circulating isoleucine level resulted in increasing 31% of peripheral atherosclerosis risk （OR=1.31， 95%CI： 1.07‒1.61）. Similarly， the OR（95%CI） was 1.33 （1.04‒1.71） in the WME model. The MR-Egger regression and MR-PRESSO analysis indicated no evidence of pleiotropy in IVs （all P>0.05）. The result of the leave-one-out sensitivity analysis was stable. The Wald ratio model displayed that the causal relationship between circulating leucine and valine levels and the risk of peripheral atherosclerosis was not statistically significant. The OR （95%CI） for leucine and valine was 1.13 （0.78‒1.63） and 1.11 （0.82‒1.50）， respectively. ConclusionThere is a significant causal relationship between the increase of circulating isoleucine level and a higher peripheral atherosclerosis risk. The causal relationships between circulating leucine and valine levels and the risk of peripheral atherosclerosis need to be further confirmed in future studies.

Value of platelet count and related scoring models in predicting the prognosis of hepatitis B virus-related acute-on-chronic liver failure / 临床肝胆病杂志

Objective To investigate the association between platelet count (PLT) and the prognosis of hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF), to establish a new PLT-related scoring model, and to assess its value in predicting the short-term prognosis of HBV-ACLF. Methods A retrospective cohort study was conducted among the patients with HBV-ACLF who were hospitalized and treated in Department of Gastroenterology, The General Hospital of Western Theater Command, from January 2018 to January 2022. Clinical data within 24 hours after admission were collected from all patients, and according to the survival after 180 days of follow-up, the patients were divided into survival group and death group. The independent-samples t test was used for comparison of normally distributed continuous data between groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups; the chi-square test was used for comparison of categorical data between groups. The Pearson correlation coefficient was used to investigate the correlation between different indicators, and the logistic regression model was used to analyze the influencing factors for prognosis. The receiver operating characteristic (ROC) curve was used to assess the predictive value of the prognostic model, and the Kaplan-Meier curve analysis was used to investigate the survival condition of the high AIP group and the low AIP group. Results A total of 236 patients were enrolled, with a 180-day survival rate of 75.85% (179/236). Compared with the survival group, the death group had significantly higher age (53.98±10.45 vs 47.44±12.46, P =0.001), international normalized ratio (INR) [1.78 (1.46-2.04) vs 1.47 (1.23-1.68), P < 0.001], total bilirubin [275.60 (165.00-451.45) vs 230.60 (154.90-323.70), P =0.035], Model for End-Stage Liver Disease (MELD) score [21.47 (18.14-24.76) vs 18.67 (15.70-21.62), P < 0.001], and albumin-bilirubin (ALBI) score [-1.06 (-1.64~-0.86) vs-1.32 (-1.73~-1.01), P =0.034], as well as significantly lower PLT [80.00 (50.00~124.50) vs 115.00 (82.00~143.00), P =0.001] and platelet-to-white blood cell ratio (PWR) [13.40 (9.54~20.70) vs 18.49 (13.95~24.74), P =0.001]. The Pearson correlation analysis showed that PLT was negatively correlated with liver cirrhosis and INR ( r =-0.332 and -0.194, P < 0.001 and P =0.003). The multivariate logistic regression analysis showed that age (odds ratio [ OR ]=1.045, 95% confidence interval [ CI ]: 1.015-1.076), PLT ( OR =0.990, 95% CI : 0.983-0.998), and INR ( OR =2.591, 95% CI : 1.363-4.925) were independent risk factors for the 180-day prognosis of HBV-ACLF patients. The new predictive model was established as follows: AIP=0.006×age+0.187×INR-0.001×PLT. The AIP scoring model had an area under the ROC curve (AUC) of 0.718 in predicting the 180-day prognosis of HBV-ACLF patients, with a sensitivity of 81.1% and a specificity of 54.1%, while PLT, PWR, LPACLF score, MELD score, and ALBI score had an AUC of 0.673, 0.659, 0.588, 0.647, and 0.578, respectively. The AIP scoring model had an optimal cut-off value of 0.48. The Kaplan-Meier survival analysis showed that the high AIP group had a significantly lower survival rate than the low AIP group ( P < 0.001). Conclusion The PLT-related scoring model has a better value than other models in predicting the prognosis of HBV-ACLF, and HBV-ACLF patients with a relatively high PLT level tend to have a high overall survival rate.

Construction of cell factories for production of valencene in Saccharomyces cerevisiae / 药学学报

Valencene, a kind of sesquiterpenoid with a citrus flavor, is mainly found in Valencia orange and is commonly used in cosmetics and food additives, as well as industrial synthetic nootkatone. In this study, synthetic biology was used to create a Saccharomyces cerevisiae cell factory to produce valencene. Fistly, valencene synthase gene (CnVS) from Callitropsis nootkatensis was inserted into the chromosome of the chassis strain YTT-T5. The resulting strain VAL-01 could produce 1.1 mg·L-1 valencene. Protein fusion technique was used, different valencene synthases were compared and the copy number of key genes was adjusted, yielding valencene to 436.4 mg·L-1. Then, knocking-out the transcription factor ROX1 resulted in valencene improvement by 17.4%. Moreover, the induction system of galactose was regulated, transcription factor PDR3 and INO2 were overexpressed. The engineered strain VAL-10 could produce 2 798.6 mg·L-1 valencene by high cell density fermentation method (nearly 2 500 times higher than VAL-01). This study provides a basis for green production of valencene.