AIM:To investigate the modulatory role of E3 ubiquitin-protein ligase ITCH in Alzheimer disease(AD)-like pathology through the thioredoxin-interacting protein(TXNIP)-nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)signaling pathway using both in vivo and in vitro experimental models.METHODS:(1)Ten 5×FAD(AD model)mice and 10 wild-type(WT)mice at 2-,4-and 6-month-old were randomly allocated into AD and WT groups.Amyloid β-protein(Aβ)plaque burden in the brain was detected by thioflavin-S and immunofluorescence staining,with the latter method additionally applied to assess TXNIP protein expression.The protein levels of ITCH and TXNIP were determined by Western blot,while their interaction was verified by co-immunoprecipitation.(2)Mouse mi-croglia BV2 cells stimulated by lipopolysaccharide(LPS)were used to construct neuroinflammation model,and were di-vided into control(CON)group and LPS+ATP treatment group.The BV2 cells stimulated by Aβ were used to construct AD inflammation model.According to the different treatment time,they were divided into CON,and 12,24 and 48 h treatment groups.Western blot was used to evaluate the expression of ITCH,TXNIP,and NLRP3 inflammasome compo-nents(NLRP3 and caspase-1)as well as the downstream IL-1β.Adenovirus-mediated ITCH overexpression(OE-ITCH)in Aβ-stimulated BV2 cells comprised three experimental groups:negative control group,Aβ oligomer stimulation group,and OE-ITCH group,with subsequent immunoblotting of inflammatory mediators.RESULTS:The deposition of Aβ plaques in the cortex and hippocampus of 5×FAD transgenic mice exhibited an age-dependent progression(P<0.01).Compared with WT mice,the levels of TXNIP protein increased synchronously,and the levels of ubiquitin ligase ITCH was significantly down-regulated(P<0.05).Co-immunoprecipitation confirmed the interaction between ITCH and TXNIP proteins in the brain of 2-and 4-month-old 5×FAD mice,which exhibited marked attenuation by 4 months of age.In BV2 microglial models,Aβ/LPS stimulation provoked significant ITCH suppression,concurrently up-regulating TXNIP,core NLRP3 inflammasome components(NLRP3 and caspase-1),and downstream IL-1β(P<0.05).Overexpression of ITCH significantly inhibited Aβ-induced activation of TXNIP and NLRP3 and therelated inflammatory factors in BV2 cells.CONCLUSION:The results of in vitro and in vivo experiments showed that ITCH protein exerts effects against AD-like pathology by inhibiting the expression of TXNIP-NLRP3 signaling pathway.

AIM:To investigate the modulatory role of E3 ubiquitin-protein ligase ITCH in Alzheimer disease(AD)-like pathology through the thioredoxin-interacting protein(TXNIP)-nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)signaling pathway using both in vivo and in vitro experimental models.METHODS:(1)Ten 5×FAD(AD model)mice and 10 wild-type(WT)mice at 2-,4-and 6-month-old were randomly allocated into AD and WT groups.Amyloid β-protein(Aβ)plaque burden in the brain was detected by thioflavin-S and immunofluorescence staining,with the latter method additionally applied to assess TXNIP protein expression.The protein levels of ITCH and TXNIP were determined by Western blot,while their interaction was verified by co-immunoprecipitation.(2)Mouse mi-croglia BV2 cells stimulated by lipopolysaccharide(LPS)were used to construct neuroinflammation model,and were di-vided into control(CON)group and LPS+ATP treatment group.The BV2 cells stimulated by Aβ were used to construct AD inflammation model.According to the different treatment time,they were divided into CON,and 12,24 and 48 h treatment groups.Western blot was used to evaluate the expression of ITCH,TXNIP,and NLRP3 inflammasome compo-nents(NLRP3 and caspase-1)as well as the downstream IL-1β.Adenovirus-mediated ITCH overexpression(OE-ITCH)in Aβ-stimulated BV2 cells comprised three experimental groups:negative control group,Aβ oligomer stimulation group,and OE-ITCH group,with subsequent immunoblotting of inflammatory mediators.RESULTS:The deposition of Aβ plaques in the cortex and hippocampus of 5×FAD transgenic mice exhibited an age-dependent progression(P<0.01).Compared with WT mice,the levels of TXNIP protein increased synchronously,and the levels of ubiquitin ligase ITCH was significantly down-regulated(P<0.05).Co-immunoprecipitation confirmed the interaction between ITCH and TXNIP proteins in the brain of 2-and 4-month-old 5×FAD mice,which exhibited marked attenuation by 4 months of age.In BV2 microglial models,Aβ/LPS stimulation provoked significant ITCH suppression,concurrently up-regulating TXNIP,core NLRP3 inflammasome components(NLRP3 and caspase-1),and downstream IL-1β(P<0.05).Overexpression of ITCH significantly inhibited Aβ-induced activation of TXNIP and NLRP3 and therelated inflammatory factors in BV2 cells.CONCLUSION:The results of in vitro and in vivo experiments showed that ITCH protein exerts effects against AD-like pathology by inhibiting the expression of TXNIP-NLRP3 signaling pathway.

Objective:To evaluate the safety and clinical efficacy of primary suturing of dog bite injuries on the head and face.Methods:A total of 42 cases with the dog bite injuries of Grade Ⅲ rabies exposure on the head and face(20 females and 22 males, aged from 1 to 81) were enrolled from October 2015 to October 2018 at the Department of Plastic Surgery, The Second People’s Hospital of Gansu Province. The primary suturing was performed after meticulous wound debridement and sufficient irrigation, the patients who were under 11 years old only received rabies vaccine for the rabies post-exposure prophylaxis, whereas the others were reinforced with tetanus antitoxin (1 500 IU) and human rabies immunoglobulin (20 IU/kg) around the wound, followed by Dermatix anti-scarring treatment for at least one year. Post-operative observation included wound healing and potential complications. All patients were followed up for facial deformities and bilateral symmetry, and the scars were assessed using the Vancouver Scar Scale (VSS).Results:Among the 42 cases, 38 cases (90.4%) were Class-A healed, 2 cases (4.8%) were Class-B healed (one of the two cases had a mandibular wound deep in the muscular layer. A deep abscess appeared in the neck 10 days after surgery, and was cured after puncture and drainage), and 2 cases (4.8%) were Class-C healed. Thirty-two patients had fever to different extent 2 to 5 days after surgery, and their body temperatures were below 38.5 ℃. The body temperature returned to normal after 1 to 2 days. During 6-24 months of follow-up, no tetanus and rabies occurred in all cases. During 1 to 2 years of follow-up, 1 of 42 cases had asymmetrical cheek 1 year post-operation. 1 case had scars on the upper eyelid 1 year after operation, and both eyes were asymmetric when eyes were open. Other 40 cases were recovered with bilateral symmetry without dysfunction of opening and closing the mouth and eyes. Facial contour recovery was cosmetically satisfactory with VSS score 1-3.Conclusions:Primary suturing of dog bite injuries on the head and face, according to the rabies prevention and treatment guideline, is a safe and effective treatment. It prevents rabies, and facial deformities and severe scar deformity in patients.

Objective:To evaluate the safety and clinical efficacy of primary suturing of dog bite injuries on the head and face.Methods:A total of 42 cases with the dog bite injuries of Grade Ⅲ rabies exposure on the head and face(20 females and 22 males, aged from 1 to 81) were enrolled from October 2015 to October 2018 at the Department of Plastic Surgery, The Second People’s Hospital of Gansu Province. The primary suturing was performed after meticulous wound debridement and sufficient irrigation, the patients who were under 11 years old only received rabies vaccine for the rabies post-exposure prophylaxis, whereas the others were reinforced with tetanus antitoxin (1 500 IU) and human rabies immunoglobulin (20 IU/kg) around the wound, followed by Dermatix anti-scarring treatment for at least one year. Post-operative observation included wound healing and potential complications. All patients were followed up for facial deformities and bilateral symmetry, and the scars were assessed using the Vancouver Scar Scale (VSS).Results:Among the 42 cases, 38 cases (90.4%) were Class-A healed, 2 cases (4.8%) were Class-B healed (one of the two cases had a mandibular wound deep in the muscular layer. A deep abscess appeared in the neck 10 days after surgery, and was cured after puncture and drainage), and 2 cases (4.8%) were Class-C healed. Thirty-two patients had fever to different extent 2 to 5 days after surgery, and their body temperatures were below 38.5 ℃. The body temperature returned to normal after 1 to 2 days. During 6-24 months of follow-up, no tetanus and rabies occurred in all cases. During 1 to 2 years of follow-up, 1 of 42 cases had asymmetrical cheek 1 year post-operation. 1 case had scars on the upper eyelid 1 year after operation, and both eyes were asymmetric when eyes were open. Other 40 cases were recovered with bilateral symmetry without dysfunction of opening and closing the mouth and eyes. Facial contour recovery was cosmetically satisfactory with VSS score 1-3.Conclusions:Primary suturing of dog bite injuries on the head and face, according to the rabies prevention and treatment guideline, is a safe and effective treatment. It prevents rabies, and facial deformities and severe scar deformity in patients.

Objective: To understand the prevalence of gynecologic diseases among married female workers. Methods: The data of married female workers who underwent occupational health examination in a physical examination center from January to December 2017 were collected. The relationship between the detection of common gynecological diseases, age and occupational types examined by gynecological routine, TCT, breast B-mode ultrasound, uterus and appendix B-mode ultrasound were analyzed. Results: Among the 1142 female workers, the total detection rate of reproductive tract infections was 67.25% (768/1142), the total detection rate of breast-related diseases was 75.22% (859/1142) ; the total detection rate of gynecological tumors and benign lesions was 14.71% (168/1142). The detection rate of breast hyperplasia was the highest 67.08% (766/1142), followed by vaginitis 51.66% (590/1142). Among the abnormalities detected in breast-related diseases, gynecological tumors and benign lesions, the highest detection rate was found in public institutions (85.66% and 27.13%), and the lowest was found in factory workers (70.24% and 7.89%). With the increase of age, the detection rate of breastrelated diseases (breast hyperplasia, breast cyst), gynecological tumors, benign lesions (uterine myoma), and Nessler's cyst abnormalities in married female workers increased (χ²_trend=7.647、21.653、107.411、53.802, P<0.05), while the detection rate of columnar epithelium of cervix decreased (χ²_trend=7.404, P<0.05). There was no significant difference in the total detection rate of reproductive tract infectious diseases (vaginitis, cervical polyps, cervical hypertrophy) among married famale workers of different ages (P<0.05) . Conclusion The common gynecological diseases of married female workers are affected by many factors such as age and occupation. Health examination and health education should be carried out regularly to reduce the incidence of gynecological diseases among female workers according to different ages and occupations.

BACKGROUND: Few studies have attempted to compare the differences in the prevalence and impact factors of hysterical tendencies (HTs) in adolescents. Thus, the aim of this study was to examine gender differences in the prevalence and impact factors of adolescents' HTs across three eastern Chinese provinces (Anhui, Jiangsu, and Zhejiang). METHODS: A multicenter, school-based, cross-sectional study was conducted in three provinces (Anhui, Jiangsu, and Zhejiang) in China in 2014. The sample included 10,131 middle-school students aged 13-18 years who were randomly selected using a multiphase, stratified, cluster sampling technique. A two-stage appraisal procedure was used to determine the adolescents' HTs. We also designed a multicenter, school-based, case control (1329 cases with 2661 control individuals) study to collect data on the common factors affecting this population using a common protocol and questionnaire. RESULTS: An overall positive rate of HTs among adolescents across the three eastern Chinese provinces studied was found at 13.1% (95% confidence interval (CI) 12.5-13.8%), at 14.5% (95% CI 13.3-15.7%) for females, and at 12.2% (95% CI 11.1-13.4%) for males. Gender-stratified, multiple conditional regression analyses revealed that superstitious beliefs pertaining to life, somatotype, teacher-student satisfaction, and family achievement orientation were significantly linked to HTs only in males, while left-behind adolescents, emotional and social adaptation, teacher-student support, family cohesion, and the Hospital Anxiety and Depression Scale - depression scores were significantly associated with female HTs only. The models indicated that of all the independent variables studied, family medical history was the strongest impact factor for both male HTs (adjusted matched odds ratio (amOR) = 2.92, 95% CI = 1.84-4.86) and female HTs (amOR = 2.74, 95% CI = 1.59-4.98). CONCLUSIONS HTs are prevalent among adolescents in the three eastern Chinese provinces studied. Gender differences in the prevalence and impact factors of HTs are significant in adolescents, and HTs seem to affect more females than males. Therefore, sex-specific intervention programs against HTs in adolescents should be considered to reduce HT prevalence in adolescents by modifying influential social, school, and family factors.
Adolescent ; Adolescent Behavior ; psychology ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Histrionic Personality Disorder ; epidemiology ; psychology ; Humans ; Male ; Prevalence ; Risk Factors ; Sex Factors ; Students ; psychology

Objective To analyze the comparation of national early warning score (NEWS), rapid emergency medicine score (REMS) and acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ) score in predicting prognosis of critically ill patients in emergency department (ED). Methods A retrospective study was conducted. Critically ill patients, aged > 16 years, hospitalized > 24 hours, and admitted to the ED of Nanhua Hospital Affiliated to South China University from January 2016 to June 2017 were enrolled. NEWS, REMS and APACHE Ⅱ score were calculated based on the worst value of each index within 24 hours after emergency admission. The primary endpoint was 28-day mortality. The relationship between the three scoring systems and the prognosis of patients was analyzed. The predictive value of three scoring systems for the prognosis of critically ill patients in ED was analyzed by receiver operating characteristic curve (ROC). Results A total of 119 emergency severe patients were enrolled in the study, and the 28-day mortality was 21.0%. The scores of NEWS, REMS and APACHE Ⅱ in the death group were significantly higher than those in the survival group (NEWS score: 9.40±3.19 vs. 5.72±2.35, REMS score: 12.64±4.46 vs. 7.97±3.28, APACHE Ⅱscore: 26.64±6.92 vs. 16.19±5.48, all P < 0.01). With the increase of NEWS, REMS and APACHE Ⅱ score, the 28-day mortality of patients gradually increased [28-day mortality of NEWS < 5, 5-6, ≥ 7 was 3.03% (1/34), 13.33% (4/34), 64.25% (20/51); 28-day mortality of REMS < 12, 12-16, ≥ 17 was 10.99% (10/91), 50.00% (11/22), 66.67% (4/6); 28-day mortality of APACHE Ⅱ < 15, 15-24, ≥ 25 was 2.33% (1/43), 15.09% (8/59), 69.57% (16/23), respectively, all P < 0.01]. The ROC curve analysis showed that the areas under the ROC curve (AUC)of NEWS, REMS and APACHE Ⅱ score for predicting the prognosis of emergency critically ill patients were 0.830 [95% confidence interval (95%CI) = 0.737-0.923], 0.782 (95%CI = 0.671-0.892) and 0.878 (95%CI = 0.800-0.956), respectively (all P = 0.000), and the accuracy of prediction was 57.4%, 48.6%, 65.4%, respectively. Conclusions The scores of NEWS, REMS and APACHE Ⅱ were useful in predicting prognosis of critically ill patients, with the highest accuracy of APACHE Ⅱ forecast, followed by NEWS, and the lowest of REMS. After comprehensive consideration of cost-effectiveness, NEWS is more reliable in ED.

β-Thalassemia is a global health issue, caused by mutations in the HBB gene. Among these mutations, HBB -28 (A>G) mutations is one of the three most common mutations in China and Southeast Asia patients with β-thalassemia. Correcting this mutation in human embryos may prevent the disease being passed onto future generations and cure anemia. Here we report the first study using base editor (BE) system to correct disease mutant in human embryos. Firstly, we produced a 293T cell line with an exogenous HBB -28 (A>G) mutant fragment for gRNAs and targeting efficiency evaluation. Then we collected primary skin fibroblast cells from a β-thalassemia patient with HBB -28 (A>G) homozygous mutation. Data showed that base editor could precisely correct HBB -28 (A>G) mutation in the patient's primary cells. To model homozygous mutation disease embryos, we constructed nuclear transfer embryos by fusing the lymphocyte or skin fibroblast cells with enucleated in vitro matured (IVM) oocytes. Notably, the gene correction efficiency was over 23.0% in these embryos by base editor. Although these embryos were still mosaic, the percentage of repaired blastomeres was over 20.0%. In addition, we found that base editor variants, with narrowed deamination window, could promote G-to-A conversion at HBB -28 site precisely in human embryos. Collectively, this study demonstrated the feasibility of curing genetic disease in human somatic cells and embryos by base editor system.
APOBEC-1 Deaminase ; genetics ; metabolism ; Base Sequence ; Blastomeres ; cytology ; metabolism ; CRISPR-Cas Systems ; Embryo, Mammalian ; metabolism ; pathology ; Female ; Fibroblasts ; metabolism ; pathology ; Gene Editing ; methods ; Gene Expression ; HEK293 Cells ; Heterozygote ; Homozygote ; Humans ; Point Mutation ; Primary Cell Culture ; Promoter Regions, Genetic ; Sequence Analysis, DNA ; beta-Globins ; genetics ; metabolism ; beta-Thalassemia ; genetics ; metabolism ; pathology ; therapy

Objective To investigate the value of CT perfusion imaging in evaluation of therapeutic effect and prognosis in radiotherapy for lung cancer.Methods Fifty-one cases of lung cancer who were unable or refused to be operated on,36 males and 15 females,aged 37 - 80,underwent CT perfusion imaging,29 of which only before radiotherapy and 22 before and after radiotherapy twice.The images were collected by cine dynamic scanning (5 mm/4 slices ) and input into the GE AW4.0workstation for data processing.The slice positions of CT imaging were determined according to the largest tumor size in CT scan.Regions of interest of tumor were drawn at the region corresponding to the original images of CT perfusion.Radiotherapy was performed after CT perfusion imaging.Relevant parameters,including blood flow ( BF),blood volume ( BV),mean transit time ( MTT),and permeability surface (PS) were calculated.The treatment response after radiotherapy was evaluated by RECIST.At 2 -4 weeks after the treatment,CT examination was conducted once more.Results The tests of the 51 patients showed that the BV was 13.6 ml·100 g-1,the BF was 129.5 ml·min-1 ·100 g-1,the MTT was 9.1 s,and the PS was 10.0 ml· min- 1· 100 g-1 before radiotherapy.The tests of the 22 of the 51 patients showed that the values of BV and BF after radiotherapy were 7.6 ml· 100 g-1 and 97.8 ml·min-1· 100 g-1,respectively,both lower than those before radiotherapy (11.2 and 108.7 ml·min-1·100g-1,respectively),however,both not significantly ( t =1.28,0.40,P ＞ 0.05 ) ; and the values of MTT and PS after radiotherapy were 8.9 s and 7.8 ml·min-1· 100 g-1,respectively,both not significantly higher than those before radiotherapy ( 7.2 s and 6.8 ml· min -1· 100 g-1,respectively,t =- 1.15,- 0.57,P ＞0.05 ).The mean area of tumor after radiotherapy was 1189.6 mm2,significantly less than that before radiotherapy ( 1920.3 mm2,t =3.98,P ＜0.05).The MTT of the SCLC patients was 12.9 s,significantly longer than that of the NSCLC patients (6.5 s,t =2.54,P ＜0.05).The MTT of the tumor with the area ≤ 10 cm2 was 11.2 s,significantly longer than that of the tumors with an area ＞ 10 cm2(5.8 s,t =2.59,P ＜ 0.05 ).The BV of the responder group was 19.2 ml· 100 g- 1,significantly higher than that of the nonresponder group (4.6 ml· 100 g - 1,t =3.62,P ＜ 0.05 ).There were not significant differences in all the perfusion characteristics between the cases with the disease-free advanced survival time ≤ 10 months and those with disease-free advanced survival time ＞ 10 months.Conclusions CT perfusion imaging helps in diagnosis and radiotherapy of lung cancer to a certain degree.

Objective To compare the immune effects of Coxsackievirus B3 (CVB3) capsid protein VP1 expressed bacterially, recombinant adenovirus rAd/VP1 and recombinant plasmid pcDNA3/VP1which express VP1 protein in mice. Methods After expressed in prokaryotic cells, VP1 protein was purified. Recombinant adenovirus rAd/VP1 and recombinant plasmid pcDNA3/VP1 were amplified and extracted. Six to 8-week-old, male BALB/c mice were divided into four groups randomly. Each group contained 18 mice. The mice of pcDNA3/VP1 group or VP1 protein group were immunized intramuscularly with three injections at three weeks apart, of recombinant plasmid pcDNA3/VP1 at a dose of 100 μg/mouse or recombinant protein VP1 at a dose of 50 μg/mouse. The mice of rAd/VP1 group were immunized intramuscularly twice at two weeks interval with rAd/VP1 at a dose of 1.2 × 107 PFU. The control group was mock-immunized with 100 μl of PBS intramuscularly. Mice were bled from the retroorbital sinus plexus every two weeks after each immunization. ELISA and micro-neutralization test were used to detect levels of CVB3-specific IgG antibody and neutralizing antibody titers in the sera of immunized mice. Three weeks after the last immunization, the cytotoxic T lymphocyte(CTL) killing activity of spleen lymphocytes was detected with CCK-8 assay. Subsequently, virus titers in the sera of immunized mice were determined by the 50% cell culture infective dose( CCID50 ) assay on HeLa cell monolayers and percentage of animals surviving were observed after lethal CVB3 attack over a period of 21 days. Results The titers of specific IgG antibody and neutralizing antibody in sera of VP1 protein immunized mice were higher than other groups( P ＜0.05 ). While CTL killing activity of spleen lymphocytes of VP1 protein immunized mice was lower than mice in rAd/VP1 group( P ＜0. 05). Virus titers in sera of VP1 protein immunized mice were lower than the mice in pcDNA3/VP1 or rAd/VP1 groups ( P ＜ 0.05 ), while survival rate was significantly higher than these two groups ( P ＜ 0.05 ).Conclusion VP1 protein induced higher level of humoral immune response and acquired obvious immune protection effects in mice. The immunizing potency of VP1 protein vaccine surpassed plasmid pcDNA3/VP1or recombinant adenovirus rAd/VP1. It appeared to be a promising candidate among the three different vaccines.