To facilitate the expression of GL 7 ACA acylase gene in a recombinant E coli , a fragment of the gene, in which the signal peptide was deleted by PCR method, was inserted into a prokaryotic expression vector, pET 28a By colony PCR method screening, a recombinant plasmid pET ACY was obtained and then transformed into the expression host BL21 (DE3) The influences of induction conditions such as IPTG concentration, the time of induction and the induction temperature on the expression of the recombinant protein were investigated Under optimal condition, the enzyme activity could reach 266 U/L Finally, the recombinant GL 7 ACA acylase can be easily isolated to a purity of about 80% by a simple anion ion exchange chromatography with enzyme activity recovery of 50%

Objective: To evaluate the outcomes of splenectomy in the treatment of relapsed/refractory autoimmune hemolytic anemia (AIHA). Methods: Retrospective analysis was performed in 30 cases with relapsed/refractory AIHA who were treated with splenectomy in our hospital. The pre- and post-operative blood routine indexes and responses were followed up. Results: Among the 30 relapsed/refractory AIHA patients, 20 were pure AIHA (including 13 patients with warm antibody AIHA, 2 with warm-cold double antibody AIHA and 5 with Coombs negative AIHA) and 10 were Evans syndrome. The short-term response was evaluated 10-14 days after operation, and the overall response rate (ORR) of short-term response was 90% [12 cases in complete response (CR), 6 cases in partial response (PR)] in 20 therapeutic evaluable cases. Among 13 patients with long-term follow-up data, except 3 patients with Evans syndrome died (2 cases were refractory to splenectomy, 1 case relapsed after surgery), the ORR of 10 patients with relapsed/refractory pure AIHA at 6 months and 12 months were 90% (9/10) and 70% (7/10), respectively, with a median follow-up of 14 (4-156) months. At the end of follow-up, 3 cases had maintained CR for more than 3 years. Conclusion The short-term response of splenectomy as a second-line treatment for relapsed/refractory AIHA is satisfactory, and long-term outcome of splenectomy is up to 70% at 1 year. Approximately one-third of patients could maintain sustained remission.

Gaming addiction， as a main manifestation of internet addiction， is characterized by a pattern of persistent or recurrent gaming behaviors. Gaming behaviors are common among adolescents and can result in marked distress or significant impairment in personal， family， social or other important areas of functioning. Recent research on health hazards of gaming addiction is relatively scattered. This article reviews the harm of adolescent game addiction to physical and mental health.

ObjectiveTo explore the relapse status based on the positive and negative syndrome scale （PANSS Scale） and related factors of schizophrenics in Shanghai communities， and to analyze the association between socio demographic characteristics， lifestyles， clinical characteristics and relapse. MethodsA dynamic cohort prospective study design was used in this study. From March 2018 to February 2019， a total of 189 schizophrenics in Xuhui， Hongkou， Changning， Jiading， Songjiang and Baoshan districts were enrolled successively. Baseline questionnaires were conducted through face-to-face interviews at baseline， which contained social demographic information， lifestyle information and clinical information. A follow-up was conducted every 2 weeks for a measurement of PANSS Scale for a total of 6 months. Relapse was assessed by a PANSS score increase of ≥25% from baseline （or an increase of 10 points or more if the baseline score was ≤40 points）. Univariate and multivariate Cox regression models were used to analyze the associations between relapse status （assessed by PANSS Scale） and socio demographic characteristics， lifestyles， and clinical characteristics， respectively. ResultsA total of 165 community schizophrenics completed baseline and follow-up surveys， with a loss to follow-up rate of about 12.7%. After exclusion of sociodemographic and clinical information deficits， 132 patients were included in the analysis totally， with an average age of 48.18±12.67 years， among whom 41.67% were male. Totally 33 patients relapsed during the 6-month follow-up period， with a relapse rate of 25.0%. After adjusting for gender， family history， age， employment， education， marital status， smoking， drinking， exercise frequency， medication compliance， insight， social function， violence history， stress recent events， adverse drug reactions and baseline scores of PANSS Scale， risk factors of relapse included the following four factors： age below 40 years （HR=4.47， 95%CI： 1.15-17.40）， primary school or below （HR=7.11， 95%CI： 1.54-32.83）， unemployed （HR=8.34， 95%CI： 1.78-38.98）， and adverse drug reactions （HR=5.02， 95%CI： 1.75-14.37）. ConclusionWe should pay attention to the risk factors such as age， education， employment and adverse drug reactions， in order to identify high-risk patients and to conduct timely interventions during the relapse management of schizophrenics in Shanghai community.

ObjectiveTo determine the factors associated with aggressive behaviors of patients with schizophrenia by gender in communities in Shanghai， and further develop and validate the prediction model. MethodsA total of 7 955 community patients with schizophrenia were investigated in Xuhui District， Hongkou District and Jiading District of Shanghai. Baseline information was collected from April 2018 and follow-up was conducted every 3 months for 6 months. Multivariate logistic regression was used to calculate the odd ratio （OR） and 95% CI， and determine the factors associated with aggressive behaviors of patients. The risk score for each patient was developed based on the β coefficient， and the best cut-off value was determined by the Youden index. For the models， predictive ability was determined using area under the curve （AUC） of receiver operator characteristic curve （ROC curve）， and internal validation ability was evaluated by the ten-fold cross validation method. ResultsThere were 3 563 males in this study with an average age of （54.83±13.72） years old， and the incidence of aggressive behaviors was 2.55%.There were 4 392 females with an average age of （57.20±14.98） years， and the incidence of aggressive behaviors was 2.64%. For male patients with schizophrenia， single/divorced status （OR=2.04， 95%CI： 1.15‒3.61）， low economic status （OR=2.79， 95%CI： 1.71‒4.54）， irregular medication （OR=4.35， 95%CI：2.23‒8.47）， no medication （OR=1.83， 95%CI：1.03‒3.26）， incomplete/no insight （OR=1.97， 95%CI：0.99‒3.94）， adverse drug reaction （OR=2.61， 95%CI：1.27‒5.37）， psychiatric symptoms involving violence （OR=2.06， 95%CI：1.01‒4.18）， history of aggression （OR=5.29， 95%CI：2.33‒11.98） and recent stress events （OR=8.36， 95%CI：4.13‒16.92） were associated with aggressive behaviors. In contrast， for female patients， age less than 60 years （50‒59 years， OR=2.09， 95%CI： 1.13‒3.87； 40‒49 years， OR=2.74， 95%CI： 1.46‒5.17； 30‒39 years，OR=2.88， 95%CI： 1.48‒5.60； 18‒29 years， OR=5.71， 95%CI： 2.44‒13.37）， educational level of high school and above （senior high school， OR=3.30， 95%CI： 1.46‒7.49； college and university， OR=2.88， 95%CI： 1.21‒6.82）， unemployed status （OR=1.81， 95%CI=1.17‒2.82）， irregular medication （OR=7.87， 95%CI：4.75‒13.05）， no medication （OR=2.11， 95%CI：1.24‒3.62）， adverse drug reaction （OR=2.75， 95%CI：1.50‒5.04）， psychiatric symptoms involving violence （OR=3.08， 95%CI：1.77‒5.37）， social function （OR=3.51， 95%CI：2.07‒5.94） and recent stress events （OR=5.92， 95%CI： 2.82‒12.44） were risk factors. In both male and female， the prediction models for aggressive behaviors of community patients with schizophrenia had strong predictive ability （AUC=0.779， 95%CI： 0.725‒0.834； AUC=0.822， 95%CI： 0.780‒0.863）. ConclusionThis study suggests that diverse risk factors should be considered for community patients with schizophrenia by gender to prevent the aggressive behaviors.

ObjectiveTo understand the stress level of people seeking psychological counseling under the coronavirus disease 2019 （COVID⁃19） pandemic and to explore its related factors. MethodsAn online survey was conducted on 1 194 people who sought psychological counseling in Shanghai through the “health cloud” psychological counseling service platform. The questionnaire included demographic information，lifestyle and stress during the COVID-19 pandemic. ResultsParticipants with low，medium，high and very high stress levels accounted for 33.1% （395/1 194），34.6% （413/1 194），25.4% （303/1 194） and 7.0% （83/1 194），respectively. Women and participants aged 18 to 30 years had higher stress levels（Z=-5.368，P<0.001； Z=35.822，P<0.001） compared with other groups. Factors contributing to the rise in stress included reading too much information about COVID-19 （OR=2.057，95%CI：1.012‒4.181），large changes in sleep state （OR=3.496，95%CI：1.669‒7.325），lack of hobbies and interests （OR=2.852，95%CI：1.252‒6.500），and prone to anxiety/irritability/sadness （OR=4.098，95%CI：1.772‒9.480）. Conclusionpeople who sought psychological counseling show high levels of psychological stress during the COVID-19 pandemic. We should pay more attention to the vulnerable groups with the following characteristics： women，18‒30 years old， residents who pay too much attention to the pandemic information，sleep less， and almost lose interest in hobbies， and easily become anxious/irritable/sad.