Objective To explore the molecular mechanism by which TP53 regulating pyroptosis,invasion and migration of embryonic fibroblasts through MMP1/NLRP3 signaling pathway.Methods NIH-3T3 murine embryonic fibroblasts were transfected with lentivirus and grouped as Control,Vector,oeTP53,oeTP53+shNC,and oeTP53+shMMP1.Cell proliferation and viability were assessed via CCK-8 assay,apoptosis by flow cytometry,and migration/invasion through scratch and invasion experiments.Protein interaction between P53 and MMP1 was confirmed by Co-immunoprecipitation.RT-qPCR evaluated mRNA expression of TP53,Collagen Ⅰ,Collagen Ⅲ,and α-SMA,while Western blot analyzed protein levels of these markers and pyroptosis-related proteins.Transmission electron microscopy was employed to examine cellular pyroptotic body modifications.Results Com-pared with Control and Vector groups,the oeTP53 group showed reduced cell proliferation activity(P<0.01),in-creased cell apoptosis rate(P<0.0001),decreased invasion(P<0.0001)and migration capabilities(P<0.0001);reduced Collagen I(P<0.001),Collagen Ⅲ(P<0.01),and α-SMA(P<0.01)protein expressions;increased NLRP3(P<0.05)and cleaved-caspase-1 expressions(P<0.01);and numerous pyroptotic bodies.MMP1 protein levels were found to be elevated in the oeTP53 group(P<0.05),and Co-IP demonstrated an interaction between p53 and MMP1 proteins.Compared with oeTP53 group,the oeTP53+shMMP1 group showed increased cell viability(P<0.001),decreased cell apoptosis rate(P<0.01),and increased cell migration(P<0.01)and invasion capabilities(P<0.01),increased scar formation-related protein expressions of Collagen I(P<0.01),Collagen III(P<0.001),and α-SMA(P<0.05);decreased pyroptosis-related protein expressions of NLRP3(P<0.01)and cleaved-caspase-1(P<0.001);and reduced pyroptotic bodies.Conclusion Overexpression of TP53 can inhibit mouse embryonic fibroblast proliferation,migration,and invasion,reduce scar formation-related protein expressions,and promote cell pyroptosis,with its mechanism potentially related to the MMP1/NLRP3 pathway.

As a new class of acid inhibitors,potassium-competitive acid blocker(P-CAB)inhibits the conformational transition of H+,K+-ATPase with subsequent suppression of H+,K+exchanging by binding reversibly near the K+binding site of H+,K+-ATPase,which results in the inhibition of gastric acid secretion in a K+-competitive manner.The unique structure and novel mechanism of P-CAB contribute to the pharmaceutical characteristics superior to other PPIs,making it a new alternative for acid-related diseases(ARDs).Progress on pharmaceutical characteristics of P-CAB were reviewed in this paper.

Objective:To explore the curative effect and prognostic assessment value of percutaneous kyphoplasty(PKP)in treating osteoporosis-caused spinal vertebral compression fractures by using imaging parameters of pelvic sagittal X-ray.Methods:A total of 198 patients with osteoporosis-caused spinal vertebral compression fractures who received treatment from January 2022 to January 2023 were selected,and they were divided into effective group(171 cases)and ineffective group(27 cases)according to the treatment effect.All patients underwent PKP treatment,and the parameters of preoperative measurements included sagittal vertical axis(SVA)of cervical vertebra 7(C7),the thoracic vertebra 1 pelvic angle(T1PA),thoracic kyphosis(TK),lumbar lordosis(LL)and sacral slope(SS).The clinically curative effect and prognosis between two groups were compared.Results:After PKP treatment,49 cases of 198 patients were cured,and 69 cases appeared significant effect,and 53 cases were effective,and 27 cases were ineffective.The SVA,LL and SS levels of effective group were significantly lower than those of ineffective group,and the differences of them were statistically significant(t=6.485,3.250,2.325,P<0.05),and the T1PA and TK of the effective group were significantly higher than those of the ineffective group(t=2.387,3.245,P<0.05),respectively.In 198 patients,39 cases occurred postoperative complications(20 cases occurred bone cement leakage,and 15 cases occurred recurrent adjacent vertebral fracture,and 2 cases occurred pulmonary embolism,and 2 cases occurred others),and 159 cases did not occurred complications.The SVA,LL and SS levels in patients without complications were significantly lower than those in patients with complications(t=10.304,5.669,0.844,P<0.05),and the T1PA and TK of patients without complications were significantly higher than those of patients with complications,and the differences were significant(t=3.494,5.550,P<0.05),respectively.The results of receiver operating characteristic(ROC)curve analysis showed that SVA,T1PA,TK,LL and SS had a certain value in assessing the curative effect and prognosis of PKP in treating osteoporosis-caused spinal vertebral compression fractures.Conclusion:Preoperative detection of imaging parameters of pelvic sagittal X-ray can assess the curative effect and prognosis of osteoporosis-caused spinal vertebral compression fractures,which can provide a certain reference for clinical treatment.

Objective:To evaluate the current status of alpha-fetoprotein (AFP) detection, a comparability analysis was conducted on the results measured by eight automated immunoassay systems, incorporating external quality assessment (EQA) data from the Beijing Center for Clinical Laboratories (BCCL) for the years 2020, 2021, and 2023.Methods:Methodological evaluation. Abbott Architect i2000, Beckman DxI 800, Roche Cobas E601, Diasorin Liaison XL, Maccura IS1200, Autolumo A2000, Leadman CI1000, and Mindray CL-2000i were used to detect 40 individual AFP serum samples that were collected from the laboratory of Beijing Chaoyang Hospital in 2019. The AFP results from eight different systems were compared with the median cohort. Passing-Bablok regression was used to evaluate the correlation between methods, and the concordance correlation coefficient was used to analyse the consistency between methods. Taking the optimal biological variability (±5.90%) as the criterion for bias evaluation, the bias between systems was evaluated using Bland-Altman analysis. The EQA results for AFP from BCCL over the past three years were statistically analysed to calculate the robust mean, robust coefficient of variation ( CV), and standard uncertainty within groups. The acceptance limit is based on the requirement of desirable biological variability (±21.87%) of allowable total error, and the pass rates were calculated for instrument or method groups, respectively. Results:The CVs of the eight detection systems were all≤1/3 allowable total error (±8.3%), passing the precision verification. The average relative biases between two detection systems (Roche Cobas E601 and Maccura IS1200) and the median cohort were>±5.90%, while the other six detection systems were<±5.90%. The eight detection systems showed good correlation and consistency with the median cohort (both R2 and concordance correlation coefficients>0.95). The results of EQA showed that there were no statistically significant differences in the robust means within each instrument or method group ( P>0.05). In the instrument group, except for Siemens and two other groups, the robust CVs of other groups were within 9%. The pass rates of most instruments and methods after being grouped were higher than the total pass rate, but that of the enzyme immunoassay chemiluminescence method was relatively low. Conclusions:The eight automated AFP immunoassay systems show a good correlation with the median cohort, and the consistency of AFP detection results is satisfactory among most detection systems. However, the comparability of AFP detection results for certain systems needs further improvement.

Ischemic stroke is one of the leading causes of death and disability worldwide. In Han dynasty， HUA Tuo proposed the original preventive medicine idea that "with good blood circulation， the disease cannot be born"， which opened a broad space for the cross-research of blood-related mechanical factors and pharmacology. In the pathogenesis of ischemic stroke， mechanical factors comprehensively affect the function and crosstalk of platelets and endothelial cells. In recent years， as the well-known effects on thrombosis and stroke， more attention has been paid to hemodynamic factors as the participants involved in pathological mechanisms and potential therapeutic targets of ischemic stroke. The mechanical force ion channel Piezo1 widely exists on the surface of many types of cells. Besides being regulated by chemical and endogenous substances， Piezo1 responds to different mechanical conditions， regulates the opening and closing of channels， and activates different downstream signaling pathways. Piezo1 is now regarded as an important connection between mechanical and biochemical signals. A variety of Chinese medicine can affect the activity of Piezo1 protein， which may prevent and treat thrombotic diseases such as ischemic stroke through Piezo1 protein. In this paper， the effects of Piezo1 protein on the physiological and pathological functions of endothelial cells and platelet under different mechanical conditions and the role of Piezo1 in the process of thrombosis were reviewed， as well as the effects of Chinese medicine， chemical medicine， and endogenous substances targeting Piezo1 channel. These could provide new ideas for further exploring the mechanisms of Chinese medicines in activating blood circulation， developing new drugs， and deepening biomechanical-pharmacology research.

Objective:To detect the mRNA expression and methylation status of leucine rich repeat containing 55(LRRC55) gene in pancreatic carcinoma tissues, and discuss the clinical value.Methods:Resected pancreatic ductal adenocarcinoma and normal adjacent specimens from 37 patients admitted in General Surgery of First Affiliated Hospital of Naval Medical University were collected from May 2019 to May 2021. Another two normal pancreas specimens and two blood samples from healthy adults were also collected. All patients′ age, gender, tumor location, tumor size, tumor differentiation, TNM staging, lymphatic metastasis, CEA and CA19-9 level were recorded. Bisulfite treatment of genomic DNA and sequencing analysis was used to study methylation patterns in CpG islands of the promoter for LRRC55 gene in fresh tissues from 2 pancreatic adenocarcinoma and adjacent tissues, 2 normal pancreatic tissues, 2 pancreatic cancer cell lines (PaTu8988 and ASPC1). LRRC55 mRNA in 35 pancreatic adenocarcinoma and adjacent tissues was detected by real-time quantitative PCR and the correlations with clinical parameters were analyzed.Results:CpG islands of LRRC55 in pancreatic adenocarcinoma tissues and pancreatic cancer cell lines was highly methylated and the mean methylation rate was 53% and 71%, respectively; while LRRC55 gene in pancreatic adjacent tissues and normal pancreatic tissues was lowly methylated, and the mean methylation rate was 8% and 11%. The relative expression in the pancreatic adenocarcinoma tissues and the paired adjacent normal tissues was 0.21 (0.02, 1.00 ) and 0.98 (0.33, 3.66 ), respectively; the former was significantly lower than the later and the difference was statistically significant ( P=0.003). Correlation analysis showed that LRRC55 mRNA expression level was related to tumor differentiation and CEA, but not correlated with patients′ age, gender, tumor location and size, CA19-9 level, lymphatic metastasis and TNM staging. Conclusions:Pancreatic cancer tissue and cell lines had abnormal methylation of LRRC55 gene; LRRC55 gene hypermethylation was related with its lower mRNA expression level in pancreatic cancer, which was correlated with the tumor differentiation and CEA level. LRRC55 may be a potential suppressor gene for pancreatic cancer.

ObjectiveBased on the inhibitory activity of phosphodiesterase （PDE）， a method for determining the anti-inflammatory activity of Qingjin Huatantang was established to supplement and improve the quality control system of this famous classical formula. MethodHigh performance liquid chromatography （HPLC） was used to determine the activity of PDE， and the dose-effect relationship of inhibiting PDE activity of Qingjin Huatantang was investigated. The mobile phase consisted of methanol-0.5% acetic acid aqueous solution （5∶95）， and the detection wavelength was 254 nm. By measuring the PDE inhibition rate of multiple batches of Qingjin Huatantang water extract lyophilized powder， biological activity was marked with the activity of the neutralizing enzyme in the international unit U. ResultWhen the concentration of reaction substrate （cyclic adenosine monophosphate） was 50 μmol·L^-1 and the reaction time was 60 min， the enzymatic reaction was stable with 4 U·mL^-1 of PDE. In this reaction system， when the concentration of Qingjin Huatantang water extract lyophilized powder was 0.11-3.0 g·L^-1， the inhibitory effect of PDE showed a concentration-dependent relationship. It was determined that the concentration of Qingjin Huatantang water extract lyophilized powder to be tested was 1 g·L^-1， which showed a significant and stable inhibitory effect on PDE， and the inhibitory rate was >45%， that is， 1 mg of Qingjin Huatantang water extract lyophilized powder could neutralize the activity of 1.8 U PDE at least. ConclusionThis study establishes a biological activity evaluation method of Qingjin Huatantang based on the inhibitory activity of PDE， and the anti-inflammatory activity of Qingjin Huatantang is characterized by international unit U of PDE activity， which can provide a new method for the determination of biological activity of traditional Chinese medicine compounds.

Objective:To establish the sex-, age-and season-specific (month) reference intervals (RI) for thyroid stimulating hormone (TSH) measurement by big data and indirect method in adults.Methods:TSH data of anonymous patients were collected from Beijing Chaoyang Hospital Affiliated to Capital Medical University in 2016, the data were selected and outliers were removed. Indirect methods (Hoffmann method and Bhattacharya method) were used to calculate TSH reference intervals of whole population, different genders, ages and seasons (months). TSH RI from two indirect methods of total population, selected population, physical examination population was compared with RI from reagent instruction according to reference change value ( RCV) based on biological variability. Results:A total of 61 599 records were obtained from 90 699 records including 18 776 males and 42 823 females. The TSH RI were obtained by Hoffmann method: the whole population, 0.59-5.59 μIU/ml (1 μIU/ml=1 mIU/L), male, 0.53-5.16 μIU/ml, female, 0.59-6.11 μIU/ml. The upper limits of TSH RI were higher with age and in winter (January): 18-30 years old, 0.62-5.57 μIU/ml, 71-80 years old, 0.49-6.45 μIU/ml; January, 0.59-6.40 μIU/ml, August, 0.60-5.56 μIU/ml; The RI of TSH by Bhattacharya method: the whole population, 0.58-5.80 μIU/ml, male, 0.55-5.02 μIU/ml, female, 0.62-6.21 μIU/ml. The upper limits of TSH RI were also higher with age and in winter (January): 18-30 years old, 0.65-5.67 μIU/ml, 71-80 years old, 0.46-5.99 μIU/ml, January: 0.61-6.52 μIU/ml, August: 0.61-5.69 μIU/ml. Compared to RI from reagent instruction, the differences of TSH RI from two indirect methods of total population, selected population, physical examination population were acceptable.Conclusions:TSH RI was established by indirect method. With the increase of age and winter, the upper limit of TSH reference interval tends to increase.

The present study was conducted to clarify the therapeutic effect of cornuside on experimental autoimmune encephalomyelitis (EAE) and its influence on T helper 17 (Th17) cell and regulatory T (Treg) cell infiltration into the central nervous system. Rats were randomly placed into four treatment groups: control, EAE, EAE+cornuside, and EAE+prednisolone. The neurological function scores of rats were assessed daily. On the second day after EAE rats began to show neurological deficit symptoms, the four groups were treated with normal saline, normal saline, cornuside (150 mg/kg), and prednisolone (5 mg/kg), respectively. The treatment was discontinued after two weeks, and the spinal cord was obtained for hematoxylin and eosin (H&E) and luxol fast blue staining, as well as retinoic acid receptor-related orphan receptor γ (RORγ) and forkhead box protein P3 (Foxp3) immunohistochemical staining. Blood was collected for Th17 and Treg cell flow cytometry testing, and the serum levels of interleukin (IL)-17A, IL-10, transforming growth factor-β (TGF-β), IL-6, IL-23, and IL-2 were measured via enzyme-linked immunosorbent assay (ELISA). Compared with rats in the EAE group, rats in the EAE+cornuside and EAE+prednisolone groups began to recover from neurological deficits earlier, and had a greater degree of improvement of symptoms. Focal inflammation, demyelination, and RORγ-positive cell infiltration were reduced by cornuside or prednisolone treatment, whereas the Foxp3-positive cell numbers were not significantly different. Meanwhile, the number of Th17 cells and the IL-17A, IL-6, and IL-23 levels were lower in the blood after cornuside or prednisolone treatment, whereas the number of Treg cells or the levels of IL-10, TGF-β, and IL-2 were not markedly different. Cornuside can alleviate symptoms of EAE neurological deficits through its anti-inflammatory and immunosuppressive effects, and Th17 cells may be one of its therapeutic targets.

Objective:To discuss the application and effectiveness of the flipped classroom based on Wechat platform in the teaching of neurology.Methods:clinical students of grade 2014 of binjiang college of Zhejiang Chinese Medical University were selected as teaching subject. The students were divided into experimental group which contained a total of 74 students in class one and class three and the control group which contained a total of 38 students in class two. Peripheral neuropathy and neuromuscular-junction disease were selected as teaching contents. The experimental group adopted flipped classroom as the teaching mode and Wechat platform for pre-class preparation, after-school review and interactive communication. The control group was taught by traditional teaching mode. Before and after class, students in the two groups had a small-scale test and were surveyed by questionnaire, respectively. Students in the two groups took the same final exam which included case analysis when the course was over. The scores of the final exam and the results of the case analysis of each group were recorded and analyzed. All data were processed with statistical software SPSS 20.0, and t-test or chi-square test was used. Results:Students in the experimental group had significantly higher test scores in the after-class small-scale test than those in the control group ( P=0.038). Their final exam scores were higher than those in the control group ( P=0.046), and their scores of case analysis in the final exam were higher than those in the control group ( P=0.026). The results of pre-class questionnaire survey showed that the proportion of students who chose "good" in the experimental group was higher than that in the control group on the understanding of the learning content and the preparation ( P<0.05). In the after-class questionnaire survey, the proportion of students who chose "excellent" and "good" in the evaluation of learning interest in the experimental group was higher than those in the control group ( P<0.05), the proportion of students in the experimental chose "good" in evaluating their self-learning ability was higher than that in the control group ( P<0.05), the proportion of "excellent" on clinical thinking ability and teaching satisfaction was higher in the experimental group than that in the control group ( P<0.05), and the overall proportion of students who chose "excellent" and "good" in all items in the experimental group were higher than that in the control group ( P<0.05). In the experimental group, the overall proportion of students who selected "excellent" and "good" in on the evaluation of their learning interest was significantly increased in the after-class questionnaire survey compared with the pre-class questionnaire survey ( P<0.01). Conclusions:The application of flipped classroom based on Wechat platform is feasible and effective in the teaching of neurology. It can make up for the deficiency of traditional teaching methods. It is helpful to improve students' learning interest and self-learning ability, and is also helpful to exercise their clinical thinking ability. Thus this method deserves further popularization.