BACKGROUND:Previously,a SENP1 gene-silenced human alveolar epithelial cell line was successfully constructed in vitro,and the role of SENP1 in hyperoxic lung injury was investigated at the cellular level.OBJECTIVE:To construct a mouse model of alveolar type II epithelial cell-specific knockout of SENP1 gene based on the Cre-loxP recombinase system.METHODS:SENP1flox/-mice were self-crossed to obtain SENP1flox/flox and SENP1flox/-mice;Sftpc-Cre+/+mice were crossed with wild-type mice to obtain more Sftpc-Cre+/-mice.Sftpc-Cre+/+or offspring Sftpc-Cre+/-mice were crossed with SENP1flox/-or offspring SENP1flox/flox mice to obtain SENP1flox/-Sftpc-Cre+/-double heterozygous mice.SENP1flox/-Sftpc-Cre+/-mice were then crossed with SENP1flox/flox mice to obtain SENP1flox/floxSftpc-Cre+/-mice.The genomic DNA was extracted by tail clipping and amplified by PCR.The amplified product was subjected to agarose gel electrophoresis to determine the mouse genotypes.Lung tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were subjected to immunofluorescence double-labelling and western blot assay to verify the knockdown effect of SENP1 gene.Heart,liver,lung and kidney tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were stained with hematoxylin-eosin to observe the histomorphology of each organ in the two groups of mice.RESULTS AND CONCLUSION:SENP1flox/floxSftpc-Cre+/-mice were correctly screened by agarose gel electrophoresis.Immunofluorescence double-labeling experiments showed that the mean fluorescence intensity of SENP1 was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with that of SENP1flox/flox mice(P<0.01)and no significant co-localization of SENP1 and Sftpc was observed(P<0.01).Western blot results showed that SENP1 protein expression was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with SENP1flox/flox mice(P<0.001).Hematoxylin-eosin staining showed no significant alterations in the histomorphology of heart,liver,lung and kidney tissues in SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice.This study successfully constructed alveolar type II epithelial cell-specific knockout SENP1 gene mice using the Cre-loxP recombinase system,which provides a good tool for the subsequent study of the role of SENP1 gene in lung diseases such as bronchopulmonary dysplasia and idiopathic pulmonary fibrosis,in which alveolar type II epithelial cells are the main damage cells.

Objective To explore the relationship between the degree of compression and shape grading of the cerebral cistern ambiens in hypertensive cerebral hemorrhage(HCH)patients after neural endoscopic intracranial hematoma evacuation(NEIHE)and prognosis.Methods 246 HCH patients who underwent NEIHE surgery from January 2020 to June 2023 were selected as the research subjects.According to the postoperative 6 month Glasgow outcome score(GOS),the patients were divided into good prognosis group(n=158)and poor prognosis group(n=88).Hierarchical regression model was used to analyze the relationship between clinical pathological features and the degree of compression and shape grading of the cerebral cistern ambiens.Multivariate logistic regression model was used to analyze the independent risk factors affecting the poor prognosis of patients with HCH after NEIHE,and a prediction model was constructed.Receiver operating characteristic curve(ROC curve)was drawn with poor prognosis as the outcome variable.Area under the curve(AUC),integrated discrimination improvement(IDI)and net reclassification improvement(NRI)of different prediction models were compared.Hosmer-Lemeshow was used to test the goodness of fit.Generalized additive model(GAM)was used to analyze the relationship between GOS and related factors.Results Compared with the good prognosis group,the poor prognosis group had more preoperative blood loss,lower preoperative Glasgow coma score(GCS),and higher proportion of patients with midline deviation,hypertension history ≥10 years,hemorrhage breaking into the brain ventricle,and operation time in the late stage,the differences were statistically significant(P＜0.05).Compared with the good prognosis group,the proportions of patients with ipsilateral and contralateral cerebral cistern ambiens compression of 3 to 4 points and the morphological classification of cerebral cistern ambiens of grade V in the poor prognosis group were higher,and the differences were statistically significant(P＜0.05).The location of bleeding,midline deviation,and intracranial pressure all had an impact on the degree of compression and morphological grading of the cerebral cistern ambiens,and the differences were statistically significant(P＜0.05).Multivariate Logistic regression analysis(model 2:including the degree of compression and shape grading of the cerebral cistern ambiens)showed that midline deviation,history of hypertension ≥10 years,preoperative bleeding volume＞37 mL,bleeding into the brain ventricles,preoperative GCS＜7 points,late surgical timing,degree of compression of the cerebral cistern ambiens(3 to 4 points on the same side of hematoma,3 to 4 points on the opposite side of hematoma),and grade V morphology were all risk factors influencing the poor prognosis of patients with HCH after NEIHE(P＜0.05).After incorporating the degree of compression and morphological grading of the cerebral cistern ambiens into model 1(excluding the degree of compression and shape grading of the cerebral cistern ambiens),the AUC increased to 0.812(95％CI:0.763～0.872),and both the IDI(0.081,95％CI:0.049～0.095)and NRI(0.611,95％CI:0.510～0.674)of the model improved,with statistically significant differences(P＜0.05).Compared with model 1,model 2 showed an increase in AUC(0.826,95％CI:0.771～0.863),IDI(0.085,95％CI:0.052～0.110),and NRI(0.628,95％CI:0.510～0.709),with statistically significant differences(P＜0.05).The Hosmer-Lemeshow test showed that model 2(P=0.878)had a better fit for predicting poor prognosis than model 1(P=0.691).GAM analysis showed that the higher the degree of compression and morphological grading of the cerebral cistern ambiens,the lower the GOS,and the difference was statistically significant(P＜0.05).ROC curve analysis showed that the combined application of degree of compression and morphological grading of the cerebral cistern ambiens had a high predictive value for poor prognosis,with AUC of 0.935(95％CI:0.890～0.971),sensitivity of 70.26％,and specificity of 93.84％.Conclusion The degree of compression and shape changes of the cerebral cistern ambiens in HCH patients after NEIHE are closely related to prognosis,and the combined application has certain predictive value for poor prognosis.The location of bleeding,midline deviation,intracranial pressure,and other factors significantly affect the compression degree and shape change of the cerebral cistern ambiens.

Objective To investigate the relationship between the levels of serum circular RNA protein tyrosine phosphatase 4A2(circ_PTP4A2),circular RNA precocious dissociation of sisters 5 homolog B(circ_PDS5B)and the volume of cerebral infarction and the degree of neurological deficits in patients with acute is-chemic stroke(AIS).Methods Ninety patients with AIS who visited the hospital from January 2021 to De-cember 2023 were selected as the AIS group,and 90 healthy individuals who underwent physical examinations during the same period were selected as the control group.The levels of serum circ_PTP4A2 and circ_PDS5B were detected by real-time fluorescence quantitative PCR,the volume of cerebral infarction was measured by magnetic resonance diffusion tensor imaging,and the degree of neurological deficit was evaluated by the Na-tional Institutes of Health Stroke Scale(NIHSS)score.According to the volume of cerebral infarction,they were divided into large-volume group(volume of cerebral infarction≥ 20 cm3,29 cases),medium-volume group(1 cm3＜volume of cerebral infarction＜20 cm3,34 cases),and small-volume group(volume of cerebral infarction≤ 1 cm3,27 cases),and according to the NIHSS scores,patients with AIS were classified into the se-vere group(NIHSS scores≥21 points,27 cases),the moderate group(NIHSS scores 5-20 points,32 cases)and the mild group(NIHSS score≤ 4 points,31 cases).Factors contributing to the increased volume of cere-bral infarction and the increased degree of neurological deficit in patients with AIS were analyzed by ordered multi-categorical Logistic regression.Results Compared with the control group,serum circ_PTP4A2 and circ_PDS5B levels were elevated in the AIS group(P＜0.05).Serum circ_PTP4A2 and circ_PDS5B levels were sequentially increased in the small-volume group,medium-volume group,and large-volume group(P＜0.05).Serum circ_PTP4A2 and circ_PDS5B levels were sequentially increased in the mild group,moderate group,and severe group(P＜0.05).Ordered multi-categorical Logistic regression showed that high NIHSS score,high circ_PTP4A2,and high circ_PDS5B were independent risk factors for increased cerebral infarction volume in AIS patients(P＜0.05),and high cerebral infarction volume,high circ_PTP4A2,and high circ_PDS5B were independent risk factors(P＜0.05).Conclusion The elevated levels of serum circ_PTP4A2 and circ_PDS5B in AIS patients are related to the increase in cerebral infarction volume and the aggravation of neurological deficits.Early detection of serum circ_PTP4A2 and circ_PDS5B levels is helpful for risk stratification in AIS patients.

Objective To systematically assess the efficacy and safety of combining acupoint massage with lactulose for constipation treatment.Methods Randomized controlled trials on the effects of acupressure combined with lactulose on constipation were searched in PubMed,Web of Science,Cochrane Library,Embase,China National Knowledge Infrastructure,SinoMed,VIP and Wanfang Data Knowledge Service Platform.The search timeframe was from the establishment of the database to July 2024,and Meta-analysis was performed using RevMan5.3 and Stata15.1 software.Results A total of 8 randomized controlled trials including 675 study participants were included.Meta-analysis showed that the acupoint massage combined with lactulose group was superior to control group in terms of total effective rate,abdominal distension score,and constipation quality of life score.Furthermore,its recurrence rate was lower than that of control group.Both groups showed no statistically significant difference in adverse reactions(P＞0.05).Conclusion Acupressure combined with lactulose can effectively improve the efficiency and quality of life in the treatment of constipation,and can be safe.Due to the limitation of the quantity and quality of the included literature,more high-quality studies with long-term follow-up are still needed to supplement and validate the above conclusions in the future.

Objective To explore the relationship between the degree of compression and shape grading of the cerebral cistern ambiens in hypertensive cerebral hemorrhage(HCH)patients after neural endoscopic intracranial hematoma evacuation(NEIHE)and prognosis.Methods 246 HCH patients who underwent NEIHE surgery from January 2020 to June 2023 were selected as the research subjects.According to the postoperative 6 month Glasgow outcome score(GOS),the patients were divided into good prognosis group(n=158)and poor prognosis group(n=88).Hierarchical regression model was used to analyze the relationship between clinical pathological features and the degree of compression and shape grading of the cerebral cistern ambiens.Multivariate logistic regression model was used to analyze the independent risk factors affecting the poor prognosis of patients with HCH after NEIHE,and a prediction model was constructed.Receiver operating characteristic curve(ROC curve)was drawn with poor prognosis as the outcome variable.Area under the curve(AUC),integrated discrimination improvement(IDI)and net reclassification improvement(NRI)of different prediction models were compared.Hosmer-Lemeshow was used to test the goodness of fit.Generalized additive model(GAM)was used to analyze the relationship between GOS and related factors.Results Compared with the good prognosis group,the poor prognosis group had more preoperative blood loss,lower preoperative Glasgow coma score(GCS),and higher proportion of patients with midline deviation,hypertension history ≥10 years,hemorrhage breaking into the brain ventricle,and operation time in the late stage,the differences were statistically significant(P＜0.05).Compared with the good prognosis group,the proportions of patients with ipsilateral and contralateral cerebral cistern ambiens compression of 3 to 4 points and the morphological classification of cerebral cistern ambiens of grade V in the poor prognosis group were higher,and the differences were statistically significant(P＜0.05).The location of bleeding,midline deviation,and intracranial pressure all had an impact on the degree of compression and morphological grading of the cerebral cistern ambiens,and the differences were statistically significant(P＜0.05).Multivariate Logistic regression analysis(model 2:including the degree of compression and shape grading of the cerebral cistern ambiens)showed that midline deviation,history of hypertension ≥10 years,preoperative bleeding volume＞37 mL,bleeding into the brain ventricles,preoperative GCS＜7 points,late surgical timing,degree of compression of the cerebral cistern ambiens(3 to 4 points on the same side of hematoma,3 to 4 points on the opposite side of hematoma),and grade V morphology were all risk factors influencing the poor prognosis of patients with HCH after NEIHE(P＜0.05).After incorporating the degree of compression and morphological grading of the cerebral cistern ambiens into model 1(excluding the degree of compression and shape grading of the cerebral cistern ambiens),the AUC increased to 0.812(95％CI:0.763～0.872),and both the IDI(0.081,95％CI:0.049～0.095)and NRI(0.611,95％CI:0.510～0.674)of the model improved,with statistically significant differences(P＜0.05).Compared with model 1,model 2 showed an increase in AUC(0.826,95％CI:0.771～0.863),IDI(0.085,95％CI:0.052～0.110),and NRI(0.628,95％CI:0.510～0.709),with statistically significant differences(P＜0.05).The Hosmer-Lemeshow test showed that model 2(P=0.878)had a better fit for predicting poor prognosis than model 1(P=0.691).GAM analysis showed that the higher the degree of compression and morphological grading of the cerebral cistern ambiens,the lower the GOS,and the difference was statistically significant(P＜0.05).ROC curve analysis showed that the combined application of degree of compression and morphological grading of the cerebral cistern ambiens had a high predictive value for poor prognosis,with AUC of 0.935(95％CI:0.890～0.971),sensitivity of 70.26％,and specificity of 93.84％.Conclusion The degree of compression and shape changes of the cerebral cistern ambiens in HCH patients after NEIHE are closely related to prognosis,and the combined application has certain predictive value for poor prognosis.The location of bleeding,midline deviation,intracranial pressure,and other factors significantly affect the compression degree and shape change of the cerebral cistern ambiens.

BACKGROUND:Previously,a SENP1 gene-silenced human alveolar epithelial cell line was successfully constructed in vitro,and the role of SENP1 in hyperoxic lung injury was investigated at the cellular level.OBJECTIVE:To construct a mouse model of alveolar type II epithelial cell-specific knockout of SENP1 gene based on the Cre-loxP recombinase system.METHODS:SENP1flox/-mice were self-crossed to obtain SENP1flox/flox and SENP1flox/-mice;Sftpc-Cre+/+mice were crossed with wild-type mice to obtain more Sftpc-Cre+/-mice.Sftpc-Cre+/+or offspring Sftpc-Cre+/-mice were crossed with SENP1flox/-or offspring SENP1flox/flox mice to obtain SENP1flox/-Sftpc-Cre+/-double heterozygous mice.SENP1flox/-Sftpc-Cre+/-mice were then crossed with SENP1flox/flox mice to obtain SENP1flox/floxSftpc-Cre+/-mice.The genomic DNA was extracted by tail clipping and amplified by PCR.The amplified product was subjected to agarose gel electrophoresis to determine the mouse genotypes.Lung tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were subjected to immunofluorescence double-labelling and western blot assay to verify the knockdown effect of SENP1 gene.Heart,liver,lung and kidney tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were stained with hematoxylin-eosin to observe the histomorphology of each organ in the two groups of mice.RESULTS AND CONCLUSION:SENP1flox/floxSftpc-Cre+/-mice were correctly screened by agarose gel electrophoresis.Immunofluorescence double-labeling experiments showed that the mean fluorescence intensity of SENP1 was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with that of SENP1flox/flox mice(P<0.01)and no significant co-localization of SENP1 and Sftpc was observed(P<0.01).Western blot results showed that SENP1 protein expression was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with SENP1flox/flox mice(P<0.001).Hematoxylin-eosin staining showed no significant alterations in the histomorphology of heart,liver,lung and kidney tissues in SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice.This study successfully constructed alveolar type II epithelial cell-specific knockout SENP1 gene mice using the Cre-loxP recombinase system,which provides a good tool for the subsequent study of the role of SENP1 gene in lung diseases such as bronchopulmonary dysplasia and idiopathic pulmonary fibrosis,in which alveolar type II epithelial cells are the main damage cells.

Objective To systematically assess the efficacy and safety of combining acupoint massage with lactulose for constipation treatment.Methods Randomized controlled trials on the effects of acupressure combined with lactulose on constipation were searched in PubMed,Web of Science,Cochrane Library,Embase,China National Knowledge Infrastructure,SinoMed,VIP and Wanfang Data Knowledge Service Platform.The search timeframe was from the establishment of the database to July 2024,and Meta-analysis was performed using RevMan5.3 and Stata15.1 software.Results A total of 8 randomized controlled trials including 675 study participants were included.Meta-analysis showed that the acupoint massage combined with lactulose group was superior to control group in terms of total effective rate,abdominal distension score,and constipation quality of life score.Furthermore,its recurrence rate was lower than that of control group.Both groups showed no statistically significant difference in adverse reactions(P＞0.05).Conclusion Acupressure combined with lactulose can effectively improve the efficiency and quality of life in the treatment of constipation,and can be safe.Due to the limitation of the quantity and quality of the included literature,more high-quality studies with long-term follow-up are still needed to supplement and validate the above conclusions in the future.

Dry eye disease(DED)refers to a condition characterized by reduced stability of the tear film or an imbalance in the microenvironment of the ocular surface, resulting from abnormalities in quality, quantity and kinetics of tear. This condition leads to various ocular discomforts and even visual impairment. The pathogenesis of DED is multifactorial and current treatment mainly focuses on symptom relief and preservation of visual function. Acupuncture has shown effectiveness in treating dry eye, although its underlying mechanism remains incompletely understood. Proteomics technology offers a comprehensive and systematic approach to studying the functions, structures and interactions of proteins. Its application in DED research can provide valuable insights into the dynamic changes in protein levels associated with different etiology or the course of DED and facilitate the identification of potential biomarkers. Furthermore, proteomics can systematically explore the regulatory mechanisms underlying acupuncture treatment for DED, providing a theoretical basis for acupuncture treatment research and contributing to the understanding of its effects at a fundamental level. This paper aims to explore the potential application of proteomics in both clinical and basic research on DED. Ultimately, it strives to offer scientific and effective strategies for the diagnosis and treatment of DED and advance our knowledge of the mechanisms underlying acupuncture therapy.

Objective To investigate the relationship between the expression of serum microRNA-326(miR-326)and microRNA-623(miR-623)in non-small cell lung cancer(NSCLC)patients and their clinical pathological characteristics and prognosis.Methods A total of 114 NSCLC patients diagnosed in our hospital from March 2019 to June 2020 were collected as study subjects as case group,123 healthy individuals who underwent physical examination were as the control group.According to the 3-year prognosis,patients were separated into a survival group of 71 cases and a death group of 43 cases.Patient related clinical data were collected,real-time fluorescence quantitative PCR method was applied to detect the expression levels of miR-326 and miR-623 in various serum samples;Kaplan-Meier method was applied to analyze the relationship between the expression levels of serum miR-326 and miR-623 in NSCLC patients and their 3-year prognosis;Cox proportional risk regression model was applied to analyze the influencing factors of 3-year prognosis in NSCLC patients.Results The expression levels of serum miR-326 in the case group and control group were 0.64±0.15 and 1.02±0.23,respectively,and the expression levels of miR-623 were 0.56±0.10 and 0.98±0.15,respectively.The difference between the two groups was statistically significant(P＜0.05).The proportions of patients with low expression of miR-326 and miR-623 in low differentiation,TNM stage Ⅲ+Ⅳ,and lymph node metastasis were higher than those in high differentiation,TNM stage Ⅰ and Ⅱ,and no lymph node metastasis(P＜0.05).The 3-year survival rates of patients with low expression of miR-326(20/55,36.36％)and miR-623(27/61,44.26％)in the serum of NSCLC patients were lower than those of patients with high expression of miR-326(51/59,86.44％)and miR-623(44/53,83.02％)(Log Rank x2=32.060,22.812,P＜0.05).Serum miR-326[(0.55±0.09)vs.(0.69±0.11)]and miR-623 levels[(0.48±0.08)vs.(0.61±0.10)]of patients in the death group were significantly lower than those in the survival group(P＜0.05).The area under the curve(AUC)for poor prognosis of serum miR-326 and miR-623 alone and in combination in patients diagnosed with NSCLC were 0.828(95％CI:0.754 to 0.901),0.763(95％CI:0.671 to 0.855),and 0.903(95％CI:0.849 to 0.958),respectively.The proportions of patients with TNM stage Ⅲ+Ⅳ,lymph node metastasis,low expression of miR-326 and low expression of miR-623in the death group were higher than those in the survival group(P＜0.05).MiR-326 and miR-623 were protective factors affecting 3-year mortality in NSCLC patients,while TNM staging and lymph node metastasis were independent risk factors affecting 3-year mortality in NSCLC patients(P＜0.05).Conclusion The low expression of miR-326 and miR-623 may be involved in the occurrence and development of lung cancer,which is closely related to the clinical pathological characteristics and poor prognosis of patients.

Objective:To observe the effects and explore the mechanism of Tuina(Chinese therapeutic massage)static training on vascular endothelial cell dysfunction and adiponectin(APN)in obese rats. Methods:Eight of 40 Sprague-Dawley rats were randomly selected as a normal group to be fed a common diet.The remaining rats were fed a high-fat diet to induce obesity models and randomly divided into a model group,an aerobic exercise group,and a static training group after successful modeling,with 8 rats in each group.During the entire experiment,rats in the normal group were fed a common diet without intervention.Rats in the model group were fed a high-fat diet without intervention.Rats in the aerobic exercise group were given moderate-intensity running exercises.Rats in the static training group were given 6-day/week static training for 8 weeks.The body mass and length of rats were recorded throughout the experiment,and the Lee's index was calculated.After the intervention,the perirenal and peri-epididymal fat weights of rats were measured,and the fat/body mass ratio was calculated.Four items of blood lipids were detected by biochemical methods[triglyceride(TG),total cholesterol(TC),low-density lipoprotein(LDL),and high-density lipoprotein(HDL)];serum nitric oxide(NO),endothelin-1(ET-1),soluble thrombomodulin(sTM),and APN were detected by enzyme-linked immunosorbent assay.The morphological changes of the aorta in each group were observed under the microscope. Results:Compared with the normal group,the body mass,body length,Lee's index,perirenal fat mass,peri-epididymal fat mass,fat/body mass ratio,TG,TC,LDL,ET-1,and sTM in the model group were significantly increased(P<0.05),while the HDL,NO,and APN were significantly reduced(P<0.01).Compared with the model group,the body mass,Lee's index,perirenal fat mass,peri-epididymal mass,fat/body mass ratio,TC,LDL,ET-1,and sTM in the aerobic exercise group and the static training group were significantly reduced(P<0.05 or P<0.01),while the HDL,NO,and APN were significantly increased(P<0.05 or P<0.01).Compared with the aerobic exercise group,NO in the static training group was significantly higher(P<0.05).APN was positively correlated with NO(P<0.01),while APN was negatively correlated with ET-1 or sTM(P<0.01).Aortic morphological observation showed that the degree of endothelial injury in the aerobic exercise group and the static training group was less significant than that in the model group,and the improvement of endothelial cell morphology in the static training group was more notable than that in the aerobic exercise group. Conclusion:Tuina static training and aerobic exercise improved the aortic endothelial injury in obese rats,and the effect of Tuina static training was more significant.The mechanism of action may be related to regulating lipid metabolism and promoting APN secretion.