Clinical management of atopic dermatitis (AD) is challenged by its susceptibility to recurrence, side effects, and high costs. We found that Portulaca oleracea L.-derived nanovesicles (PDNV) exert anti-inflammatory effects by modulating macrophage M1/M2 polarization. These effects were achieved through pathways including inhibition of nuclear factor-κB (NF-κB) and stimulator of interferon genes (STING) protein expression in diseased tissues, demonstrating their potential to ameliorate AD symptoms. To increase the transdermal permeation of PDNV, dissolvable microneedles composed primarily of hyaluronic acid (HA) were developed as an adjunctive means of delivery. Meanwhile, polysaccharides of Portulaca oleracea L., which were synergistic with PDNV, were used as microneedle constituent materials to enhance the mechanical properties and physical stability of HA. This new means of delivery significantly improves the treatment of AD and also provides new options for the efficient utilization of plant extracellular vesicles and the treatment of AD. In addition, transcriptomic analysis of PDNV showed that the mRNAs of Portulaca oleracea L. are closest to those of ferns, which may shed light on related evolutionary and plant species identification studies.

Objective:To evaluate the effect of esketamine on cerebral ischemia-reperfusion (I/R) injury and the association with mitochondrial stress in mice.Methods:The experiment was performed in two parts. Part Ⅰ Eighteen SPF male C57BL/6 mice, aged 8-12 weeks, with body mass index of 28-30 g, were divided into 3 groups ( n=6 each) by a random number table method: sham operation group (S group), cerebral I/R group (IR group), and esketamine plus cerebral I/R group (E+ IR group). Cerebral I/R was produced by occlusion of middle cerebral artery for 1 h followed by 24-h reperfusion in anesthetized mice.Esketamine 10 mg/kg was intraperitoneally injected at 20 min before developing the model in E group. Neurological function was evaluated using the Zea Longa score and balance beam test (Feeney score). The cerebral infarct size was determined by TTC staining. Part Ⅱ Primary cortical neurons were isolated and cultured and then divided into 3 groups ( n=42 each) using a random number table method: control group (group C), oxygen-glucose deprivation-reoxygenation (OGD/R) group, and esketamine plus OGD/R group (group E+ OGD/R). Cells were subjected to O 2-glucose deprivation for 1 h followed by restoration of O 2-glucose supply for 24 h. The cells were treated with 25 μmol/L esketamine for 40 min before preparing the model in E+ OGD/R group. The neuronal viability was measured by the CCK-8 assay. The ultrastructure of neurons was observed with a transmission electron microscope. The levels of reactive oxygen species (ROS), glutathione peroxidase (GSH-px) and malondialdehyde (MDA) were determined, and the mitochondrial membrane potential was determined by JC-1 kit. The neuronal apoptosis was detected by TUNEL staining, and the apoptosis rate of neurons was calculated. The expression of Bax, cytochrome C (CytC), cleaved-caspase-9, caspase-3 and cleaved-caspase-3 was detected by Western blot. Results:Part Ⅰ Compared with S group, the Zea Longa score, Feeney score and cerebral infarct size were significantly increased in IR group ( P<0.01). Compared with IR group, the Zea Longa score, Feeney score and cerebral infarct size were significantly decreased in E+ IR group ( P<0.01). Part Ⅱ Compared with C group, the cell viability and activity of GSH-px were significantly decreased, the apoptosis rate of neurons, levels of ROS and MDA, mitochondrial membrane potential, and cleaved-caspase-3/caspase-3 ratio were increased, and the expression of Bax, Cyt C and cleaved-caspase-9 was up-regulated in OGD/R group ( P<0.01). Compared with OGD/R group, the cell viability and activity of GSH-px were significantly increased, the apoptosis rate of neurons, levels of ROS and MDA, mitochondrial membrane potential, and cleaved-caspase-3/caspase-3 ratio were decreased, and the expression of Bax, Cyt C and cleaved-caspase-9 was down-regulated in E+ OGD/R group ( P<0.01). Conclusions:Esketamine can alleviate cerebral I/R injury in mice, and the mechanism may be related to inhibition of mitochondrial stress in neurons, improvement in mitochondrial function, and inhibition of mitochondria-dependent apoptosis in neurons.

ObjectiveTo screen out the transcriptomes related to the intervention of Wuzi Yanzongwan on the spermatogenic function of semi-castrated male mice， and to explore its potential mechanism in the intervention of the progress of low spermatogenic function. MethodBalb/c mice were randomly divided into sham-operated group， model group， testosterone propionate group（0.2 mg·kg^-1·d^-1， intramuscular injection） and Wuzi Yanzongwan group（1.56 g·kg^-1·d^-1， intragastric administration） according to body weight， with 12 mice in each group. The right testicle and epididymis were extracted from the model group and the drug administration group to construct the semi-castrated model of low spermatogenic function， while the fur and the right scrotum of the sham-operated group were only cut and immediately sterilized and sutured. At the end of the intervention， hematoxylin-eosin（HE） staining was used to observe the histopathology of testis， enzyme-linked immunosorbent assay（ELISA） was used to detect the levels of serum testosterone（T）， luteinizing hormone（LH） and follicle stimulating hormone（FSH）. The sperm count and motility of epididymis were measured by automatic sperm detector of small animal. Transcriptomic microarray technology was used to detect the mRNA expression level of testicular tissue in each group， the transcriptome of genes related to the regulation of Wuzi Yanzongwan was screened， and three mRNAs were selected for Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） to verify the transcriptome data. Through the annotation analysis of Gene Ontology（GO） and the signaling pathway analysis of Kyoto Encyclopedia of Genes and Genomes（KEGG）， the related functions of drugs regulating transcriptome were analyzed. ResultCompared with the sham-operated group， the testicular tissue of mice in the model group showed spermatogenic injury， contraction and vacuolization of the seminiferous tubules， reduction of spermatogenic cells at all levels， widening of the interstitial space， obstruction of spermatogonial cell development and other morphological abnormalities， and serum T significantly decreased， LH significantly increased（P<0.01）， and FSH elevated but no statistically significant difference， the count and vitality of epididymal sperm significantly decreased（P<0.01）. There were 882 differentially expressed mRNAs in the testicular tissues， of which 565 were up-regulated and 317 were down-regulated. Cluster analysis showed that these differentially expressed mRNA could effectively distinguish between the sham-operated group and the model group. Compared with the model group， the damage to testicular tissue in the Wuzi Yanzongwan group was reduced， the structure of the seminiferous tubules was intact， vacuolization was reduced， and the number of spermatogenic cells at all levels was significantly increased and arranged tightly. The serum T significantly increased， LH significantly decreased（P<0.01）， and FSH decreased but the difference was not statistically significant. The count and vitality of sperm in the epididymis were significantly increased（P<0.01）. Moreover， Wuzi Yanzongwan could regulate 159 mRNA levels in the testes of semi-castrated mice， of which 32 were up-regulated and 127 were down-regulated， and the data of the transcriptome assay was verified to be reliable by Real-time PCR. GO and KEGG analysis showed that the transcriptome functions regulated by Wuzi Yanzongwan were involved in the whole cell cycle process of sperm development such as sex hormone production of interstitial cells in testis， renewal， differentiation， metabolism， apoptosis and signal transduction of spermatogenic cells， and were closely related to the biological behaviors of signaling pathways such as spermatogenic stem cell function， endoplasmic reticulum protein processing and metabolic program. ConclusionWuzi Yanzongwan can effectively improve the low spermatogenic function of semi-castrated male mice， and its mechanism may be related to the regulation of testicular transcriptional regulatory network， the synthesis of sex hormones in testicular interstitial cells， the function of spermatogenic stem cells， the whole cell cycle process of spermatogenesis， as well as the expression of endoplasmic reticulum protein processing and metabolic program related genes transcription.

Objective:To evaluate the role of autophagy in morphine preconditioning-induced reduction of oxygen-glucose deprivation and restoration (OGD/R) injury in primary cortical neurons of mice and the relationship with c-Jun N-terminal kinase (JNK).Methods:Primary cortical neurons extracted from C57BL/6 neonatal mice within 24 h after birth were divided into 9 groups ( n=24 each) using a random number table method: control group (C group), OGD/R group, morphine preconditioning group (M group), autophagy inhibitor 3-methyladenine (3-MA) group (3-MA group), 3-MA+ morphine preconditioning group (3-MA+ M group), autophagy inhibitor chloroquine group (Ch group), chloroquine + morphine preconditioning group (Ch+ M group), JNK inhibitor SP600125 group (SP group) and SP600125 + morphine preconditioning group (SP+ M group). Morphine preconditioning: morphine was added at a final concentration of 3 μmol/L before OGD/R, and the cells were incubated for 2 h in OGD/R group. In 3-MA, Ch and SP groups, 3-MA 5 mmol/L, chloroquine 50 μmol/L and SP600125 25 μmol/L were added, respectively, and the cells were incubated for 150 min. In 3-MA+ M, Ch+ M and SP+ M groups, 3-MA 5 mmol/L, chloroquine 50 μmol/L and SP600125 25 μmol/L were added, respectively, at 30 min before morphine preconditioning. Then the cells were subjected to oxygen-glucose deprivation for 1 h followed by restoration of oxygen-glucose supply for 24 h. CCK-8 assay was used to detect the neuronal viability. The expression of JNK, phosphorylated JNK (p-JNK), microtubule-associated protein 1 light chain 3 (LC3), p62, Beclin1, caspase-3, and cleaved-caspase-3 was determined by Western blot. The autophagosomes and autolysosomes were counted using LC3-double fluorescent adenovirus transfection, and the neuronal apoptosis rate was determined by TUNEL staining. Results:Compared with C group, the neuronal viability was significantly decreased, the expression of Beclin1 was up-regulated, the expression of p62 was down-regulated, and the LC3Ⅱ/LC3Ⅰ ratio, p-JNK/JNK ratio, the number of autophagosomes and autolysosomes, cleaved-caspase-3/caspase-3 ratio and neuronal apoptosis rate were increased in OGD/R group ( P<0.001). Compared with OGD/R group, the neuronal viability, p-JNK/JNK ratio, LC3Ⅱ/LC3Ⅰ ratio and the number of autophagosomes and autolysosomes were significantly increased, the expression of Beclin1 was up-regulated, and the expression of p62 was down-regulated in M group, the LC3Ⅱ/LC3Ⅰ ratio was significantly decreased, and the expression of p62 was down-regulated in 3-MA group, the LC3Ⅱ/LC3Ⅰ ratio was significantly increased, and the expression of p62 was up-regulated in Ch group ( P<0.001), and no significant change was found in the parameters mentioned above in SP600125 group ( P>0.05). Compared with M group, the neuronal viability was significantly decreased, the LC3Ⅱ/LC3Ⅰ ratio was decreased, and the expression of p62 was up-regulated in M+ 3-MA group, the neuronal viability was significantly decreased, the LC3Ⅱ/LC3Ⅰ ratio was increased, and the expression of p62 was up-regulated in M+ Ch group, and the neuronal viability, LC3Ⅱ/LC3Ⅰ ratio and p-JNK/JNK ratio were significantly decreased, the expression of p62 was up-regulated, the number of autophagosomes and autolysosomes was decreased, the expression of Beclin1 was down-regulated, and the cleaved-caspase-3/caspase-3 ratio and neuronal apoptosis rate were increased in M+ SP group ( P<0.001). Conclusions:Morphine preconditioning can attenuate OGD/R injury by activating JNK, enhancing autophagy and inhibiting apoptosis in primary cortical neurons of mice.

Objective:To investigate the clinical features of IgD multiple myeloma (MM) and the effect and prognosis of daratumumab-based combination therapy.Methods:The clinicopathological data of a IgD MM patient with disease progression and extramedullary infiltration treated with daratumumab in the Affiliated Hospital of Qingdao University in December 2019 were retrospectively analyzed.Results:The 74-year-old woman was diagnosed as IgD MM by bone marrow aspiration and immunofixation electrophoresis. The patient was given VD (bortezomib, dexamethasone), RD (lenalidomide, dexamethasone) and ID (ixazomib, dexamethasone) regimens. In June 2020, the patient developed multiple subcutaneous nodules, and she was assessed as progressive disease with extensive extramedullary infiltration. After treated with daratumumab-PAD (liposomal doxorubicin, bortezomib, dexamethasone) regimen, the patient's subcutaneous nodules were significantly reduced and partially disappeared, and the general condition was significantly improved. But the patient was in a cachexia state and finally died of the irregular treatment and disease progression.Conclusions:IgD MM has a low incidence and a short survival period, and there is no uniform standard treatment. The early application of daratumumab combined with proteasome inhibitors, immunomodulators, cytotoxic drugs and hematopoietic stem cell transplantation may improve the overall survival of patients.

Objective: To understand the status of e-cigarette use among junior high school students in Zhengzhou City and its associated factors, and to provide support for tobacco control intervention for junior high school students. Methods: From March to May 2019 in Zhengzhou, a stratified cluster random sampling method was used to investigate 3 624 junior high school students using a self-made e-cigarette questionnaire, including demographic information, attitudes towards e-cigarettes, usage behavior, smoking behaviors of parents and friends. Results: The usage rate of e-cigarettes for junior high school students in Zhengzhou was 5.5%, parents and 51.5% of junior high school students purchased e-cigarettes online, 28.3% purchased them through stores, and 20.2% purchased e-cigarettes through other methods. Regarding the item use of e-cigarettes was harmful to others, the proportion of students with an approved attitude was lower than that with negative attitudes, the difference was statistically significant(χ2=4.33, P<0.05). Logistic regression analysis showed that e-cigarette was difficult to quit, the use of e-cigarettes won t make people feel relaxed, e-cigarette smoke was harmful to others , e-cigarette use did not make young people more attractive were negatively correlated with the use of e-cigarettes by middle school students(OR=0.46, 0.28, 0.57, 0.41, P<0.01). Conclusion The use of e-cigarettes by junior high school students in Zhengzhou is relatively serious, and multiple measures should be taken to strengthen health education and reduce the use rate of e-cigarettes.

Objective: To investigate the effect of immunophenotyping on prognosis of multiple myeloma (MM) patients treated with bortezomib regimen as main treatment. Methods: Seventy-six MM patients in the Department of Hematology in the Affiliated Hospital of Qingdao University from January 2012 to January 2017 were retrospectively analyzed. The effects of the expressions of CD45, CD56 and other factors on progression free survival (PFS) and overall survival (OS) in MM patients treated with bortezomib-containing regimen were also analyzed. Results: Univariate analysis showed that statistical differences of the median PFS (12 months vs. 19 months, P < 0.001) and median OS (19 months vs. 23 months, P= 0.001) were significant in the group of CD45 positive and negative of MM patients; the median PFS (14 months vs. 21 months, P= 0.014) and median OS (16 months vs. 25 months, P= 0.026) for MM patients with hemoglobin (Hb) < 100 g/L and Hb ≥ 100 g/L were statistically significant; the median PFS (13 months vs. 18 months, P= 0.004) and median OS (14 months vs. 24 months, P= 0.008) for MM patients with albumin (ALB) < 35 g/L and ALB ≥ 35 g/L were statistically significant. The median PFS time and the median OS time in female MM patients were shorter than those in male MM patients (13 months vs. 19 months, P= 0.008; 16 months vs. 25 months, P= 0.008). Multivariate analysis showed that female and CD45 positive were the independent poor prognostic factors for PFS and OS in MM patients (P < 0.05). Conclusion CD45 positive and female are important prognostic factors for MM patients treated with bortezomib regimen as main treatment.

Objective To investigate prognostic factors in patients with primary central nervous system diffuse large B-cell lymphoma (PCNS-DLCBL).Methods The clinical data of 71 patients with PCNS-DLCBL confirmed by pathology and clinical tests, who were admitted to our hospital from 1991 to 2015, were retrospectively analyzed.All patients received chemotherapy, mainly with high-dose methotrexate (HD-MTX, 66/71), and 59 patients received radiotherapy, mainly with whole brain radiotherapy (WBRT)±local boost.The Kaplan-Meier method was used to calculate survival rates, the log-rank test was used for survival comparison and univariate prognostic analysis, and the Cox model was used for multivariate prognostic analysis.Results Fifty-eight patients achieved a complete response (CR), ten achieved a partial response (PR), and three had progressive disease (PD).The 5-year overall survival (OS) rate was 43%, and the 5-year progression-free survival (PFS) rate was 34%.The univariate analysis showed that the factors associated with OS included the following:age of onset, Karnofsky Performance Scale (KPS) score, single or multiple lesions, whether to receive radiotherapy, evaluation results after radiotherapy and chemotherapy, and the presence or absence of recurrence (P=0.000-0.047);the multivariate analysis showed that the three factors that affected OS were age of onset, KPS score, and the presence or absence of recurrence (P=0.000-0.022).The univariate analysis revealed that chemotherapy regimen, whether to receive radiotherapy, total radiotherapy dose, WBRT dose, evaluation results after radiotherapy and chemotherapy, and the presence or absence of recurrence were the factors associated with PFS (P=0.000-0.028);the multivariate analysis revealed that KPS score and the presence or absence of recurrence were associated with PFS (P=0.000-0.011).Conclusions Among patients with PCNS-DLCBL, younger age, higher KPS score, and no recurrence are associated with better OS, and single lesion, radiotherapy, and better evaluation results after radiotherapy and chemotherapy may be associated with better OS;higher KPS score, better evaluation results after radiotherapy and chemotherapy, and no recurrence are the factors associated with better PFS, and HD-MTX chemotherapy, radiotherapy, higher total radiotherapy dose, and higher WBRT dose may be associated with better PFS.Whether to receive radiotherapy after achieving a CR with chemotherapy and the target area and dose of radiotherapy need to be further studied.

Objective To investigate the impact of eotaxin-3 gene polymorphisms on the clinical effect of inhaled corticosteroids (ICS) to provide clinical basis for eotaxin-3 as the target spot for treating bronchial asthma.Methods One hundred and ninety-six cases of asthma and 196 cases as controls were selected from the outpatients and inpatients in our hospital.Peripheral blood samples were collected from the asthma patients and normal controls.PCR-RFLP was adopted to detect the genotypes of eotaxin-3 +2497T＞G and-+-77C＞T.The response of ICS treatment and the change situation of ACT scores were compared among asthmatic patients with various genotypes.Results Peripheral blood eosinophil(EOS) counts,EOS proportion and total IgE in the patients with TG genotype at+2497 locus were significantly decreased compared with those in the patients with TT genotype,the difference was statistically significant(P＜0.05).The level of PD20 in asthmatic patients with TG genotype was significantly higher than that in the patients with TT genotype,the difference was statistically significant[(0.07-±-0.03)mg vs.(0.03 ± 0.01)mg,t=2.45,P=0.048];whereas the above indicators had no statistical difference among 3 kinds of +-77 genotypes.During ICS treatment process in the patients with TT genotype at +-2497 locus,the FEV1％,PD20 value and ACT scores were significantly improved compared with those in the patients with TG genotype,the difference was statistically significant(P＜0.01).Conclusion The asthmatic patients with TT genotype at +-2497 locus were more sensitive to ICS treatment,regular ICS treatment can significantly improve the lung function and clinical symptom score in these patients.