Objective:To evaluate the relationship between the mechanism of mild hypothermia-induced reduction of neuronal apoptosis during cerebral ischemia-reperfusion (I/R) and calcium/calmodulin-dependent protein kinase kinase 2 (CaMKK2)/adenosine monophosphate-activated protein kinase (AMPK) signaling pathway in rats.Methods:Forty SPF male Sprague-Dawley rats, aged 6-8 weeks, weighing 200-250 g, were divided into 5 groups ( n=8 each) using a random number table method: sham operation group (Sham group), cerebral I/R group (IR group), hypothermia + cerebral I/R group (HIR group), hypothermia + cerebral ischemia-reperfusion + solvent group (HIR-DMSO group) and hypothermia + ischemia-reperfusion + CaMKK2 inhibitor STO-609 group (HIR-STO609 group). A global cerebral I/R injury model was established using the four-vessel occlusion method. In HIR group, HIR-DMSO group, and HIR-STO609 group, an ice blanket was used to reduce the body temperature immediately after cerebral ischemia, bringing the core body temperature down to 32.5-33.5 ℃, and rewarming was carried out 4 h later. One hour before developing the model, STO-609 solution 4 μl was injected into the lateral ventricle in HIR-STO609 group, and the equal volume of dimethyl sulfoxide solution was given instead in HIR-DMSO group. At the end of reperfusion, the modified neurological severity score (mNSS) was performed. Then the rats were sacrificed under deep anesthesia, and the hippocampal tissues were taken to observe the pathological results of the hippocampal tissues (using HE staining and Nissl staining) and to determine the apoptosis rate of neurons (by TUNEL method) and expression of B-cell lymphoma 2(Bcl-2), Bcl-2-associated X (Bax), CaMKK2, phosphorylated AMPK (p-AMPK) and AMPK (by Western blot). Results:Compared with Sham group, the mNSS and apoptosis rate of neurons in the hippocampal tissues were significantly increased, the expression of Bax was up-regulated, the expression of Bcl-2, CaMKK2 and p-AMPK was down-regulated, and the ratio of p-AMPK/AMPK was decreased in IR group ( P<0.05). Compared with IR group, the mNSS and apoptosis rate of neurons in the hippocampal tissues were significantly decreased, the expression of Bax was down-regulated, the expression of Bcl-2, CaMKK2 and p-AMPK was up-regulated, and the ratio of p-AMPK/AMPK was increased in HIR group ( P<0.05). Compared with HIR group, the mNSS and apoptosis rate of neurons in the hippocampal tissues were significantly increased, the expression of Bax was up-regulated, the expression of Bcl-2, CaMKK2 and p-AMPK was down-regulated, the ratio of p-AMPK/AMPK was decreased ( P<0.05), and no significant change was found in the aforementioned parameters in HIR-DMSO group ( P>0.05). Conclusions:Mild hypothermia can inhibit neuronal apoptosis by up-regulating the CaMKK2/AMPK signaling pathway, thus reducing cerebral I/R injury in rats.

BACKGROUND:At present,chemotherapeutic drugs are mainly used for the treatment of tumors,but there are problems such as drug resistance and adverse reactions.The exosome drug delivery system not only avoids the toxicity of synthetic nanoparticles,but also increases the bioavailability and biocompatibility of the drugs.It can be modified by biological,physical,and chemical methods to form a new type of nano-drug delivery platform.OBJECTIVE:To review the construction strategy of exosome drug delivery system,the application status of exosome drug delivery system in tumor diseases and the current challenges.METHODS:PubMed and CNKI were searched with"exosomal,tumor,microvesicle,extracellular vesicles,engineered,therapeutics,characterization,isolation,drug delivery,targeting,modification strategies,physics,chemistry,biology"as English search terms and"exosomes,drug delivery,tumor"as Chinese search terms.A total of 132 articles were included for in-depth induction and discussion.RESULTS AND CONCLUSION:(1)The technical methods of exosome extraction,including ultracentrifugation,filtration,and kit extraction,can efficiently isolate exosomes,but the process is complicated and time-consuming,and large-scale extraction of exosomes cannot be achieved.(2)Engineered exosomes can be divided into four categories:gene editing engineering,which improves function through genetic modification;endogenous engineering,using inflammatory factors and other pretreatment to enhance drug delivery;exogenously engineered to encapsulate drugs directly in exosomes;hybrid engineering,combining exosomes with lipid nanoparticles to form new particles.Some have entered clinical trials for cancer treatment,but most are at an early stage.In contrast,genetically engineered exosomes are considered as an important direction for future drug delivery due to their high targeting and customization potential.(3)There are still many limitations to realize the clinical transformation of engineered exosomes.At the technical level,large-scale production,purification,and drug loading efficiency are urgent to be solved.In production,high cost and batch stability affect its popularity.In terms of safety,immunogenicity and potential toxicity need to be comprehensively evaluated.Furthermore,the imperfect regulatory policies and the complexity of the approval process also constitute obstacles to its clinical translation.(4)In the future,it is necessary to promote the clinical translation process through technical innovation,cost control,safety improvement,and policy improvement.

Objective:To evaluate the relationship between the mechanism of mild hypothermia-induced reduction of neuronal apoptosis during cerebral ischemia-reperfusion (I/R) and calcium/calmodulin-dependent protein kinase kinase 2 (CaMKK2)/adenosine monophosphate-activated protein kinase (AMPK) signaling pathway in rats.Methods:Forty SPF male Sprague-Dawley rats, aged 6-8 weeks, weighing 200-250 g, were divided into 5 groups ( n=8 each) using a random number table method: sham operation group (Sham group), cerebral I/R group (IR group), hypothermia + cerebral I/R group (HIR group), hypothermia + cerebral ischemia-reperfusion + solvent group (HIR-DMSO group) and hypothermia + ischemia-reperfusion + CaMKK2 inhibitor STO-609 group (HIR-STO609 group). A global cerebral I/R injury model was established using the four-vessel occlusion method. In HIR group, HIR-DMSO group, and HIR-STO609 group, an ice blanket was used to reduce the body temperature immediately after cerebral ischemia, bringing the core body temperature down to 32.5-33.5 ℃, and rewarming was carried out 4 h later. One hour before developing the model, STO-609 solution 4 μl was injected into the lateral ventricle in HIR-STO609 group, and the equal volume of dimethyl sulfoxide solution was given instead in HIR-DMSO group. At the end of reperfusion, the modified neurological severity score (mNSS) was performed. Then the rats were sacrificed under deep anesthesia, and the hippocampal tissues were taken to observe the pathological results of the hippocampal tissues (using HE staining and Nissl staining) and to determine the apoptosis rate of neurons (by TUNEL method) and expression of B-cell lymphoma 2(Bcl-2), Bcl-2-associated X (Bax), CaMKK2, phosphorylated AMPK (p-AMPK) and AMPK (by Western blot). Results:Compared with Sham group, the mNSS and apoptosis rate of neurons in the hippocampal tissues were significantly increased, the expression of Bax was up-regulated, the expression of Bcl-2, CaMKK2 and p-AMPK was down-regulated, and the ratio of p-AMPK/AMPK was decreased in IR group ( P<0.05). Compared with IR group, the mNSS and apoptosis rate of neurons in the hippocampal tissues were significantly decreased, the expression of Bax was down-regulated, the expression of Bcl-2, CaMKK2 and p-AMPK was up-regulated, and the ratio of p-AMPK/AMPK was increased in HIR group ( P<0.05). Compared with HIR group, the mNSS and apoptosis rate of neurons in the hippocampal tissues were significantly increased, the expression of Bax was up-regulated, the expression of Bcl-2, CaMKK2 and p-AMPK was down-regulated, the ratio of p-AMPK/AMPK was decreased ( P<0.05), and no significant change was found in the aforementioned parameters in HIR-DMSO group ( P>0.05). Conclusions:Mild hypothermia can inhibit neuronal apoptosis by up-regulating the CaMKK2/AMPK signaling pathway, thus reducing cerebral I/R injury in rats.

BACKGROUND:At present,chemotherapeutic drugs are mainly used for the treatment of tumors,but there are problems such as drug resistance and adverse reactions.The exosome drug delivery system not only avoids the toxicity of synthetic nanoparticles,but also increases the bioavailability and biocompatibility of the drugs.It can be modified by biological,physical,and chemical methods to form a new type of nano-drug delivery platform.OBJECTIVE:To review the construction strategy of exosome drug delivery system,the application status of exosome drug delivery system in tumor diseases and the current challenges.METHODS:PubMed and CNKI were searched with"exosomal,tumor,microvesicle,extracellular vesicles,engineered,therapeutics,characterization,isolation,drug delivery,targeting,modification strategies,physics,chemistry,biology"as English search terms and"exosomes,drug delivery,tumor"as Chinese search terms.A total of 132 articles were included for in-depth induction and discussion.RESULTS AND CONCLUSION:(1)The technical methods of exosome extraction,including ultracentrifugation,filtration,and kit extraction,can efficiently isolate exosomes,but the process is complicated and time-consuming,and large-scale extraction of exosomes cannot be achieved.(2)Engineered exosomes can be divided into four categories:gene editing engineering,which improves function through genetic modification;endogenous engineering,using inflammatory factors and other pretreatment to enhance drug delivery;exogenously engineered to encapsulate drugs directly in exosomes;hybrid engineering,combining exosomes with lipid nanoparticles to form new particles.Some have entered clinical trials for cancer treatment,but most are at an early stage.In contrast,genetically engineered exosomes are considered as an important direction for future drug delivery due to their high targeting and customization potential.(3)There are still many limitations to realize the clinical transformation of engineered exosomes.At the technical level,large-scale production,purification,and drug loading efficiency are urgent to be solved.In production,high cost and batch stability affect its popularity.In terms of safety,immunogenicity and potential toxicity need to be comprehensively evaluated.Furthermore,the imperfect regulatory policies and the complexity of the approval process also constitute obstacles to its clinical translation.(4)In the future,it is necessary to promote the clinical translation process through technical innovation,cost control,safety improvement,and policy improvement.

Objective:To enhance the understanding of hereditary protein C deficiency.Methods:A case of a child with severe hereditary protein C deficiency, presenting with recurrent muscle bleeding as the primary clinical manifestation, was reported. The diagnostic and treatment were detailed, and relevant literature was reviewed.Results:The patient, a 2-year-old girl, was admitted with "recurrent muscle hematomas for more than 2 years and intracranial hemorrhage for 3 months." Ecchymosis was observed on the skin of the left calf, with localized warmth detected on palpation. Muscle strength and tone in both lower limbs were within normal limits. The left thigh circumference measured 35.6 cm, compared to 29 cm on the right. Laboratory tests showed decreased fibrinogen, coagulation factor XIII activity, protein C activity, and significantly elevated fibrin degradation products (FDP) and D-dimer levels. Genetic testing identified compound heterozygous mutations in the PROC gene: c.565 (exon 7) C>T and c.983_988 (exon 7) del GCGAGC. The patient was diagnosed with hereditary protein C deficiency and disseminated intravascular coagulation (DIC). Treatment with fibrinogen supplementation, fresh frozen plasma, and anticoagulation therapy led to clinical improvement.Conclusion:The clinical manifestations of hereditary protein C deficiency are highly heterogeneous. While neonatal purpura fulminans is a common presentation, recurrent bleeding can also serve as a primary clinical manifestation.

Objective:To explore the effects of body mass index (BMI) on the incidence of nocturnal hypertension in patients with hypertension.Methods:Totally 341 hospitalized hypertensive patients treated at the Affiliated Hospital of Jining Medical University from February to May 2023 were selected by convenience sampling. Patients' general information, clinical data, and 24-hour ambulatory blood pressure results were collected. A binomial Logistic regression analysis was conducted to investigate the factors affecting the occurrence of nocturnal hypertension in these patients. The relationship between BMI and the incidence of nocturnal hypertension was examined using threshold effect tests and smooth curve fitting.Results:The binomial Logistic regression analysis indicated that blood phosphate level was a factor influencing the occurrence of nocturnal hypertension in hypertensive patients ( P<0.05). Smooth curve fitting and threshold effect test results showed that the relationship between BMI and the incidence of nocturnal hypertension was curve-correlated, with a turning point at 24.61 kg/m 2. To the left of the turning point, there was no correlation ( P=0.130) ; to the right, there was a correlation ( P=0.016) . Conclusions:When the BMI of hypertensive patients exceeds 24.61 kg/m 2, the likelihood of nocturnal hypertension increases with rising BMI, providing a precise intervention target for weight management-based patient care in hypertension.

Objective:To explore the application effect of portable device of airway humidification on patients with head and neck tumor who underwent tracheostomy.Methods:A total of 100 patients with head and neck tumors who underwent tracheostomy at Department of Head and Neck Surgery of Cancer Hospital of Chinese Academy of Medical Sciences and Peking Union Medical College from October 2021 to May 2023 were selected as the study subjects.They were randomly divided into the control group and the observation group by using a random number table method,with 50 cases in each group.The control group received conventional nebulization inhalation for airway humidification,and the observation group adopted portable device of airway humidification with 0.45%sodium chloride solution to conduct sustained airway humidification.The effect of airway humidification,occurrence of phlegm scabs,injury and bleeding of airway,tracheal tube obstruction,and comfort level of patients after surgery were observed and compared between the two groups.Results:The satisfaction rates of the patients of observation group at the 1st,2nd,3rd,4th,and 5th day after surgery were 84.0%,88.0%,92.0%,96.0%and 96.0%for humidification of wet sputum,which were better than those of control group,and the differences were statistically significant(x2=46.240,64.103,70.560,67.919,74.227,P<0.05),respectively.The formations of phlegm scabs were respectively 1,5,1 and 1 cases at the 2nd,3rd,4th,5th d after surgery in observation group,which were less than those in control group,and the differences were statistically significant(x2=7.111,23.926,21.760,17.344,P<0.05),respectively.The numbers of airway injury bleeding and overall tracheal tube obstruction at the 2nd d after surgery in the observation group were respectively 2 cases and 1 case,which were significantly less than those in the control group,and the differences were statistically significant(x2=4.000,5.982,P<0.05),respectively.The comfort levels of airway humidification and suction phlegm,and overall comfort evaluation were respectively(17.96±0.75)scores,(16.06±1.05)scores and(34.04±1.63)scores in the observation group,which all were lower than those in the control group,and the differences were statistically significant(t=9.843,17.298,15.792,P<0.05),respectively.Conclusion:The portable device of airway humidification has a significant effect on airway humidification of patients with head and neck tumors who undergo tracheostomy,which can reduce the occurrence of phlegm scabs,and reduce the number of overall tracheal tube obstruction,and improve comfort level of patient with head and neck tumors who undergo tracheostomy.

Objective:To investigate the value of dual-detector spectral CTA in distinguishing infarct core from penumbra in patients with acute ischemic stroke(AIS), and to further explore the risk factors associated with infarct core and their predictive value.Methods:The imaging and clinical data of 163 patients with AIS who met the inclusion criteria admitted to the Second Affiliated Hospital of Soochow University from March 2022 to May 2023 were retrospectively analyzed. Patients from March 2022 to December 2022 were used as the training group, and patients from January 2023 to May 2023 were used as the validation group for internal validation. The head and neck spectral CTA and brain CT perfusion imaging with dual-layer detector spectral CT were all carried out on all patients. Using CTP as reference, the patients were divided into infarct core group and non-infarct core group according to whether an infarct core occurred in the hypoperfusion regions of brain tissue. Multivariate logistic regression analysis was used to screen predictors related to the infarct core. The receiver operating characteristic (ROC) curve was used to evaluate the predictive efficacy.Results:A total of 163 patients were included in the study, including 112 in the training group and 51 in the validation group. There were significant differences in iodine density, effective atomic number, hypertension, triglyceride and neutrophils between the two groups ( P< 0.05). The cutoff values for iodine density values and effective atomic number values were 0.215 mg/mL and 7.405, respectively. Multivariate logistic regression analysis showed that iodine density and hypertension were independent risk factors for infarct core in AIS, and triglyceride was an independent protective factor. The area under the ROC curve (AUC) of iodine density value was the largest (0.859), with a sensitivity of 70.27%, and a specificity of 90.67%, which had a good predictive value. The ROC curve analysis results for the validation group were consistent with the training group. Conclusions:Spectral CT parameters iodine density values and effective atomic number values have the potential to distinguish the infarct core area from the penumbra area in patients with AIS. Iodine density and hypertension were independent risk factors of infarct core in AIS, triglyceride was an independent protective factor, and iodine density values obtained by dual-layer spectral detector CT had a high predictive value.

Objective:To discuss the effect of oridonin on the proliferation,migration,epithelial-mesenchymal transition(EMT),and apoptosis of the human nasopharyngeal carcinoma HONE-1 cells,and to clarify its related antitumor mechanism.Methods:The HONE-1 cells were treated with different concentrations(0,5,10,20,40,80,and 160 mg·L-1)of oridonin for 48 h.CCK-8 method was used to detect the inhibitory rates of proliferation of the cells in various groups and the drug concentration for subsequent experiment was confirmed.The HONE-1 cells were divided into control group,3 mg·L-1 oridonin group,and 6 mg·L-1 oridonin group.After 24 and 48 h of culture,CCK-8 method was used to detect the proliferation activities of the cells in various groups;5-ethynyl-2'-deoxyuridine(EdU)method was used to detect the rates of EdU-positive cells in various groups;colony formation assay was used to detect the numbers of clone formation in the cells in various groups;Transwell chamber experiment and cell wound assay were used to detect the numbers of migration cells and the scratch healing rates of the cells in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of cyclin-dependent kinase 1(CDK1)and cyclin-dependent kinase 4(CDK4)mRNA in the cells in various groups;Western blotting method was used to detect the expression levels of E-cadherin,Vimentin,Caspase-3,and poly ADP-ribose polymerase 1(PARP1)proteins in the cells in various groups.Results:The CCK-8 method results showed that the half-maximal inhibitory concentration(IC50)of oridonin at 48 h was 12.18 mg·L-1,and 1/4 IC50 and 1/2 IC50 values were used as the concentrations for subsequent experiments.Compared with control group,after treated for 24 and 48 h,the proliferation activities of the cells in 3 and 6 mg·L-1 oridonin groups were decreased(P＜0.05 or P＜0.01),the rate of EdU-positive cells were decreased(P＜0.05 or P＜0.01),the numbers of clone formation and migraton cells were decreased(P＜0.05 or P＜0.01),the scratch healing rates were decreased(P＜0.05 or P＜0.01),the expression levels of CDK1 and CDK4 mRNA in the cells were decreased(P＜0.05 or P＜0.01),the expression levels of E-cadherin,Caspase-3,and PARP1 proteins were increased(P＜0.05 or P＜0.01),and the expression levels of Vimentin protein were decreased(P＜0.05).Conclusion:Oridonin can inhibit the proliferation,clone formation,and migration of the human nasopharyngeal carcinoma HONE-1 cells by downregulating the expression of cell cycle-related proteins and EMT,and promote the apoptosis to exert an antitumor effect.

Background Schizophrenia patients often face high level of stigma and low level of self-esteem,significantly hindering their recovery.Mindfulness-based intervention has proven be effective in reducing stigma and improving self-esteem.However,traditional mindfulness intervention typically involve high costs and require long-term professional involvement.Objective To explore the effects of blended mindfulness interventions on stigma and self-esteem in patients with stable schizophrenia,so as to provide references for reducing stigma,enhancing self-esteem and promoting recovery.Methods Patients receiving outpatient treatment at the Third Hospital of Daqing from June 2022 to January 2023,who met the diagnostic criteria for schizophrenia in the International Classification of Diseases,tenth edition(ICD-10)and were in a stable phase,were recruited for the study(n=84).According to the random number table method,participants were randomly assigned to study group and control group,with 42 cases in each group.Both groups received treatment with the second-generation antipsychotic medications,while the study group additionally received blended mindfulness intervention for 8 weeks,with sessions lasting 45～60 minutes,three times a week.Both groups were evaluated with Five Facet Mindfulness Questionnaire(FFMQ),Internalized Stigma of Mental Illness Inventory(ISMI)and Rosenberg Self-Esteem Scale(RSES)at baseline and after 8-week intervention.Covariance analysis was used to compare the FFMQ,ISMI and SES scores between two groups after 8-week intervention.Results After 8-week intervention,there were statistically significant differences between two groups in total FFMQ scores,as well as in the observation,acting with awareness,non-judgment and non-reactivity subscale scores(F=50.680,12.952,13.567,22.799,14.043,P<0.01).Statistically significant differences were observed in total ISMI scores,and in the alienation,stereotype endorsement,discrimination experience,stigma resistance and social withdrawal subscale scores(F=513.125,148.990,125.055,75.996,154.850,54.125,P<0.01).The difference in RSES scores between two groups was also statistically significant(F=19.478,P<0.01).Conclusion Blended mindfulness intervention may help improve the mindfulness and self-esteem in stable schizophrenia patients while reducing stigma.