Objective To construct the eukaryotic expression plasmid for the expression of human Connexin26 in COS-7 cells.Methods Total RNA was isolated from human peripheral blood lymphocytes and used as template for the PCR cloning of the human Connexin26 gene.The human Cx26 cDNA containing the 678 bp whole coding region of the human Connexin26 gene was amplified by PCR using specific primers and cloned into the pCI-neo vector to construct the recombinant eukaryotic expression plasmid,pCI-Cx26.The recombinant plasmid was identified by restriction endonuclease digestion,and transfected into COS-7 cells by liposome.The expression of Cx26 mRNA and the protein were analyzed by RT-PCR and SDS-PAGE,respectively.Results Restriction endonuclease digestion analysis verified successful construction of the recombinant plasmid,pCI-Cx26.The expression of Cx26 mRNA and protein in the transfected COS-7 cells were detected by RT-PCR and SDS-PAGE,respectively.Conclusion The eukaryotic expression plasmid for human Cx26 has been constructed successfully with the capability of expression in COS-7 cells.

OBJECTIVE:To study the improvement effects of citalopram combined with huperzine A in aged depression model rats. METHODS:Aged rats were randomly divided into blank control group,model group,huperzine A group(0.3 mg/kg),citalo-pram group(5 mg/kg),and combination group(huperzine A 0.3 mg/kg+citalopram 5 mg/kg),10 in each group. Except for blank control group,rats in other groups received chronic unpredictable mild stress to reduce depression model. After modeling,rats were intragastrically administrated relevant drugs once a day,for 2 weeks. The depression,learning and memory behavior changes of rats in each group were observed by using open-field test,sucrose consumption test,tail suspension test,forced swimming test and Morris water maze test. RESULTS:Compared with blank control group,the horizontal crossing number,uprightness number,su-crose preference rate,crossing number in platform,percentages of target quadrant distance and time of rats in model group were ob-viously decreased (P<0.05 or P<0.01);immobility time of tail suspension and swimming,escape latency were obviously pro-longed(P<0.05 or P<0.01). Compared with model group,the depression-related indexes of rats in citalopram group and combina-tion group were obviously improved(P<0.05 or P<0.01),and combination group had better effects;the learning and memory-re-lated indexes in combination group were obviously improved(P<0.05 or P<0.01),only crossing time in platform in huperzine A group and citalopram group were obviously increased (P<0.05 or P<0.01),and other learning and memory-related indexes had no obvious changes(P>0.05). CONCLUSIONS:Citalopram combined with huperzine A can obviously improve the depression be-havior,learning and memory ability of aged rats with depression,showing better effects than citalopram alone.

Objective To study the time extended for getting emergency intervention in different modes of transportation and factors influencing the modes of transportation of patients with ST elevation myocardial infarction (STEMI).Methods A total of 564 consecutive patients with STEMI admitted from September 2013 to June 2016 were enrolled in the study.The clinical data about time consumed for getting emergency intervention and modes of transportation were collected.Results According to the mode of transportation,patients were divided into three groups:emergency care system (EMS) transportation group (n =96),self-transportation group (n =206) and referral group in which the patients were sent in from other hospitals (n =262).EMS transportation group had significantly shorter total ischemic time before emergency treatment than self-transportation group (229 rin vs.418 min,P ＜ 0.05) and referral group (229 min vs.512 rin,P ＜ 0.05),and significantly shorter length of pre-hospital time than self-arrival group (55 min vs.110 min;P＜0.05) and referral group (55 min vs.372 min;P＜0,05).The referral group had longer pre-hospital time and the self-transportation group had longer door-to-balloon time,but there was no difference in total ischemic time between the self-arrival and referral group (Z =-1.882,P =0.068).Multivariate logistic regression was used to analyze influence factors in mode of transportation:(1) patients characterized with high school or university education,profession of civil service,and their transportation distance more than 30 km were greater in number than referral group (P ＜ 0.05);(2) patients identified with senior middle school education,staff member of public sectors or company,their transportation distance less than 30 km,and with killip grade above Ⅱ were more likely to have EMS transport (P ＜ 0.05);(3) patients defined as businessmen without taking out new rural cooperative medical insurance,taking up transportation distance less than 80 km,and subjecting to killip grade Ⅰ had a higher proportion of individuals of this kind taking self-transportation (P ＜ 0.05).Conclusion Mode of transportation is an important factor that affects the time extended to get emergency intervention.Education level,occupation,medical insurance type,transportation distance,killip grade are associated with modes of transport.

Objective: To detect the expression level of early and late protein of vaccinia virus and to preliminarily explore replication-defective mechanism of highly attenuated NTV strain of vaccinia virus Tiantan. Methods: We constructed prokaryotic expression vector, expressed and purified homologous early protein E3 and late protein A27 closely related to replication and prepared mouse polyclonal antiserum by immunizing mice with homologous proteins. Early and late protein expression levels of NTV were detected. Results: We have expressed and purified vaccinia virus proteins respectively in E. coli expression system and prepared homologous mouse polyclonal antiserum. Early protein E3 and late protein A27 could be highly efficient expression in NTV infected non-permissive Hela cells, while expression of late protein F17 was blocked detected by Western blot. Conclusions The expression limitation of late protein F17 may be an explanation for the replication-defective mechanism of NTV.

Objective To investigate the delay of door to signature time in primary percutaneous coronary intervention (PCI) and its influence in patients with ST segment elevation myocardial infarction (STEMI),therefore to provide a scientific basis for further effective shortening the time of primary PCI in patients with STEMI.Methods A total of 226 patients who diagnosed with STEMI and underwent primary PCI at Henan Provincial People's Hospital from June 2016 to December 2017 were enrolled in the study.Observation indicators include:(1) baseline data of patients;(2) time segments in primary PCI:total ischemic time (TIT),door to balloon time (DTBT),door-to-signature time (DTST),signature to balloon time (STBT);(3) the demographic characteristics of the family members who signed informed consent;and (4) the psychological factors and coping strategies of family members before signing informed consent.All data was analyzed using SPSS software (version 22.0).Multiple linear regression analysis was used to analyze the influencing factors of delay of DTST.A P＜0.05 was considered statistically significant.Results In this study,226 patients with STEMI who were first diagnosed in our hospital had a mean age of 55.23±10.80 years,and 181 (80.1％) were male.The median of TIT,DTBT,DTST,STBT were 312 min,166 min,82 min,and 80 min.The ratio of DTST in DTBT and TIT was 50％ and 28.5％,respectively.The multiple linear regression analysis showed that the number of direct family members (P＜0.001),the degree of educational in middle school and below (P=0.010),high school/technical secondary school (P=0.029),families worrying about the high cost of medical care (P=0.020),families consulted each other repeatedly (P=0.022),and consulted the other medical staff(P=0.022) are risk factors of DTST delay,and city residence (P=0.048) is the protection factor of DTST delay.Conclusions The long time of DTS is a reality of the practice of primary PCI in China.The factors that lead to longer DTST include demographic characteristics,psychological factors and coping strategies of family members.The STBT of primary PCI in China should be taken into the value while emphasizing the DTBT.

Objective To investigate the efficacy and safety of gastric peroral endoscopic myotomy ( G-POEM) for postsurgical gastroparesis. Methods A retrospective analysis was performed on data of 47 patients with gastroparesis after subtotal gastrectomy, who underwent G-POEM at Zhongda Hospital Southeast University form August 2016 to June 2018. G-POEM procedure, complications related to G-POEM, and improvement of symptoms and gastric emptying function were observed. Results All patients underwent G-POEM successfully. No serious G-POEM-related complications were observed. During a follow-up period of 9. 8±5. 8 months (range 3-18 months), the scores of gastroparesis cardinal symptoms index (GCSI) were 1. 6±0. 5, 1. 2±0. 4, 1. 1±0. 6, 1. 4±0. 3 and 1. 7±0. 3 respectively at the 1st, 3rd, 6th, 12th, and 18th months after G-POEM, and all scores were lower than that before G-POEM (3. 8±1. 1, all P<0. 01). The results showed the symptoms of gastroparesis were improved significantly. Gastric emptying imaging showed the hemi-emptying time was 23. 8±8. 8 min, 21. 2±9. 5 min, 20. 9±8. 3 min and 26. 4±7. 8 min at the 1st, 6th, 12th and 18th months after G-POEM, respectively, significantly shorter than that before G-POEM (67. 8±12. 5 min, all P<0. 01). Three-dimensional ultrasonography results of gastric antrum volume showed that the hemi-emptying time was 26. 4 (21. 8, 40. 3) min, 22. 6 (13. 9, 32. 7) min, 24. 3 (18. 2, 36. 5) min and 26. 8 (16. 4, 38. 5) min at the 1st, 6th, 12th and 18th months after G-POEM, respectively, which were all significantly shorter than that before G-POEM [ 72. 5 ( 48. 3, 108. 6 ) min, all P<0. 01 ] . Conclusion G-POEM has a satisfactory long-term efficacy on the treatment of gastroparesis after subtotal gastrectomy with good safety.

Objective: To generate monkeypox virus specific monoclonal antibodies for further establishing monkeypox virus immunofluorescence assay. Methods: Monkeypox virus A29 protein, vaccinia ortholog A27 protein and cowpox ortholog 162 protein were expressed in E. coli BL21 to screen antibodies. Synthetic monkeypox virus A29_{17 ~ 49} polypeptide was used to immune BALB/c mice. Monkeypox virus monoclonal antibodies were generated through fusion, cloning and screening techniques. Indirect ELISA was performed to test antibodies specificity and subtype. Results: A29, A27 and 162 proteins were highly expressed in E. coli and detected by Western blot. The three his-tagged proteins were purified using His-Bind affinity chromatography column. The purity of the proteins was all more than 90%. And 8 strains monkeypox virus specific monoclonal antibodies were screened by the three purified proteins. Two mAbs of 8 were IgG3 subtype and the rest were IgG1 subtype. Conclusions Eight strains of monkeypox virus specific monoclonal antibodies were generated, they can be used to further establish monkeypox virus immune immunofluorescence assay.

This review summarizes dietary characteristics of patients with psoriasis, discusses effects of gluten-free diet, Mediterranean diet and dietary intervention-induced weight loss on psoriasis, and analyzes the efficacy of dietary supplements in the treatment of psoriasis, such as fish oil, vitamin D, vitamin B12, selenium, and probiotics.

ObjectiveTo explore the activating effects of ten important effective components in seven medicinal and edible substances on human pregnane X receptor （PXR）， including Glycyrrhizae Radix et Rhizoma （liquiritin and glycyrrhizic acid）， Houttuyniae Herba （quercetin and houttuyfonate）， Prunellae Spica （rosmarinic acid）， Cassiae Semen （aurantio-obtusin）， Poria （pachymic acid）， Lilii Bulbus （Lilium brownii saponin and colchicine）， and Lycii Fructus （Lycium barbarum polysaccharide） and screen potentially toxic components. MethodCell counting kit-8 （CCK-8） assay was used to investigate the cytotoxic effect of liquiritin， glycyrrhizic acid， quercetin， houttuyfonate， rosmarinic acid， pachymic acid， aurantio-obtusin， and colchicine （10， 20， and 50 μmol·L^-1）， and L. brownii saponin and L. barbarum polysaccharide （10， 20， and 50 mg·L^-1） on normal human hepatocyte cell line （L02）. The release of lactate dehydrogenase （LDH） in L02 cells after drug treatments was detected by the biochemical analyzer. The apoptosis induced by ten effective components was explored by Hoechst 33342 staining. The secreted luciferase reporter system was used to co-transfect the PXR expression vector and reporter gene vector containing cytochrome P450 3A4 （CYP3A4） transcriptional regulatory region into L02 cells， with 10 μmol·L^-1 rifampicin （RIF） as a positive control. After treated with liquiritin， glycyrrhizic acid， quercetin， houttuyfonate， rosmarinic acid， aurantio-obtusin， pachymic acid， and colchicine （5， 10， and 20 μmol·L^-1） and L. brownii saponin and L. barbarum polysaccharide （5， 10， and 20 mg·L^-1） for 24 h， the cells were tested for secreted luciferase activity. ResultCompared with the control group， colchicine， L. brownii saponin， and quercetin decreased the cell viability （P<0.05， P<0.01）. Compared with the control group， quercetin， rosmarinic acid， glycyrrhizic acid， colchicine， aurantio-obtusin， and pachymic acid increased the release rate of LDH in L02 cells （P<0.05， P<0.01）. The proportion of hyperchromatic nuclei increased gradually after rosmarinic acid， liquiritin， and L. barbarum polysaccharide treatments as compared with the control group （P<0.05， P<0.01）. In terms of co-transfection of pcDNA3.1-PXR and pGLuc-CYP3A4 into L02 cells， compared with the control group， aurantio-obtusin and pachymic acid showed activating effects on PXR （P<0.05）， whereas liquiritin and glycyrrhizic acid showed inhibitory effects （P<0.05）. ConclusionThe findings suggest that when medicinal and edible substances are taken for a long time， attention should be paid to their influence on drug-metabolizing enzymes and possible interactions， so as to improve their safety.

ObjectiveTo explore the mental health status and its influencing factors among middle school students in Xide County， Liangshan Yi Autonomous Prefecture， and to provide references for mental health interventions for local middle school students. MethodsUsing a cross-sectional study design， one junior middle school and one senior middle school in Xide County， Liangshan Prefecture， Sichuan Province， were randomly selected on September 16， 2021， and two classes per grade in each school involving 288 students were recruited. Subjects were assessed using Patients' Health Questionnaire Depression Scale-9 item （PHQ-9）， Generalized Anxiety Disorder Scale-7 item （GAD-7）， PTSD Checklist for DSM-5 （PCL-5）， Multidimensional Scale of Perceived Social Support （MSPSS） and UCLA Loneliness Scale （ULS-3）. Then the scores of above scales were compared among middle school students with different demographic characteristics， and binary Logistic regression analysis was conducted to screen the influencing factors of post-traumatic stress disorder (PTSD） symptoms. ResultsAmong the respondents， 17.71% （95% CI： 0.133~0.221）， 8.68% （95% CI： 0.054~0.120）， 2.78% （95% CI： 0.009~0.047） and 45.83% （95% CI： 0.400~0.516） were reported to have symptoms of depression， anxiety， PTSD and loneliness， respectively. Students in senior middle school scored lower on PCL-5 and ULS-3 than those in junior middle school ［（6.46±8.91） vs. （8.87±9.42）， t=2.202， P<0.05； （4.67±1.65） vs. （5.60±1.88）， t=4.431， P<0.01］. Regression analysis denoted that depressive symptoms （OR=7.630， P<0.05） and anxiety symptoms （OR=3.789， P<0.01） were risk factors for PTSD symptoms among middle school students. ConclusionThe middle school students in Xide County， Liangshan Yi Autonomous Prefecture suffer a high prevalence rate of depressive symptoms and loneliness， and those in junior middle school are more likely to feel a sense of strong loneliness， furthermore， depressive symptoms and anxiety symptoms are risk factors for PTSD symptoms.