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.

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.

Objective: To understand the awareness and service utilization of HIV preexposure prophylaxis (PrEP) and postexposure prophylaxis (PEP) among the young students aged 15-24 in Hefei, so as to provide reference for the prevention and treatment of AIDS among young students in the future. Methods: From May to June 2022, a stratified random cluster sampling method was used to select two different types of universities(undergraduate and vocational colleges) in Hefei, and a crosssectional survey was conducted among the 916 young students at the selected schools. The Chisquare test was used for statistical analysis. Results: Among the surveyed young students, 88.4% had heterosexual orientation, and 12.4% had sexual behavior including 8.8% of homosexual sexual behavior and 91.2% of heterosexual sexual behavior. The rate of condom use was 70.0% in the last homosexual intercourse, and the rate of condom use was 83.7% in the last heterosexual intercourse. About 41.9% of young students heard of PrEP, and the proportions of young students who knew about PrEP, consulted PrEP and used PrEP were 43.0%, 15.9% and 9.4% respectively. About 38.3% of young students had heard of PEP, and the proportions of young students who knew about PEP, consulted PEP and used PEP were 18.2%, 15.4% and 6.0%. Conclusions The awareness rate and utilization rate of PrEP and PEP among young students in Hefei are relatively low. PrEP and PEP publicity should be combined with education of HIV knowledge, and enhance the awareness rate of the young students, in order to reduce the risk of AIDS infection among young students with highrisk behaviors.

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.

Objective:To investigate the incidence and influencing factors of allergic reactions to cephalosporins.Methods:A cross-sectional study of 29 medical institutions in Zhejiang Province was conducted from April 2021 to June 2021.The incidence of allergic reactions to cephalosporins was investigated,and the influencing factors of cephalosporin-induced allergic reactions were analyzed by Poisson regression.Results:A total of 56 155 patients were included in this study.The total incidence of allergic reactions to cephalosporin was 1.67‰,the highest incidence of anaphylaxis occurred for ceftizoxime(4.27‰),followed by ceftriaxone(3.49‰)and cefotaxime(2.40‰).There was no significant difference in the incidence of allergic reactions between patients with negative skin tests and those without skin tests(1.75％o vs.1.63‰,RR=1.07,95％CI:0.70-1.63,P＞0.05).Poisson regression showed that body mass index(BMI)＜18.5 kg/m2(RR=2.43,95％CI:1.23-4.82,P＜0.05)and history of β-lactam antibiotics allergy(RR=33.88,95％CI:1.47-781.12,P＜0.05)increased cephalosporin-induced anaphylaxis.Compared with cefuroxime,the risk of allergic reactions was increased for ceftriaxone(RR=3.08,95％CI:1.70-5.59,P＜0.01),ceftazidime(RR=1.89,95％CI:1.03-3.47,P＜0.05),and ceftizoxime(RR=3.74,95％CI:1.64-8.50,P＜0.01).Conclusions:Lower BMI and history of β-lactam antibiotics allergy increase the risk of cephalosporin allergic reactions.The routine skin test may not reduce the occurrence of allergic reactions to cephalosporins. This study has been registered at Chinese Clinical Trial Registry(ChiCTR2200064314).

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.