Solamargine improves the therapeutic efficacy of anti-PD-L1 in lung adenocarcinoma by inhibiting STAT1 activation.

OBJECTIVE: The effect of solamargine on lung adenocarcinoma and its effect on STAT1 signaling pathway mediated immune escape were studied through network pharmacology and in vitro and in vivo experiments. METHODS: The solamargine targets were screened using the TCMSP and the LUAD targets were screened using the GeneCard, OMIM, PharmGkb, TTD and DrugBank databases. PPI network analysis and target prediction were performed using GO and KEGG. Colony formation assay, EDU staining, wound healing, transwell assay, Hoechst and flow cytometry were used to detect the effects of solamargine on the proliferation, migration and apoptosis of LUAD. Western blotting (WB) and quantitative reverse transcription polymerase chain reaction (RT-qPCR) were used to detect P-STAT1 and PD-L1 expression. And immunofluorescence was used to detect P-STAT1 expression. In vivo experiments, C57BL/6 mice were divided into control group, low concentration group, high concentration group, positive control group and combination group. Every other day, following seven consecutive doses, the size of the tumor was assessed. Finally, the expressions of P-STAT1, STAT1, PD-L1 and apoptosis index proteins were detected by WB. RESULTS: The anti-LUAD effect of solamargine was found by wound healing, colony formation assay, transwell assay, hoechst and EdU staining. The results of network pharmacological analysis showed that solamargine could suppress STAT1 expression level. Further enrichment assay of STAT1 showed that STAT1 was associated with immune-related pathways. In addition, molecular signal analysis by WB and RT-qPCR indicated that solamargine could reduce the expression levels of P-STAT1 and PD-L1 in a concentration-dependent manner. According to the results of in vivo assays, combination of solamargine and immune checkpoint inhibitors (ICIs) durvalumab could significantly inhibit the growth of Lewis transplanted tumors in C57BL/6 mice, and no toxic side effect was recoded. CONCLUSION: These results indicated that solamargine could inhibit the proliferation and promote the apoptosis of LUAD. It also could reduce the expression level of P-STAT1 protein and inhibit the expression level of PD-L1. At the same time, the combination with the ICIs can better block the expression of PD-L1 in cells, thereby inhibiting the immune escape pathway of tumor cells and achieving anti-tumor effects. This study proposed a novel combined therapeutic approach, involving the inhibition of STAT1 by solamargine in conjunction with ICIs.

Combinatorial optimization of CO2 transport and fixation to improve succinate production by promoter engineering.

To balance the flux of an engineered metabolic pathway to achieve high yield of target product is a major challenge in metabolic engineering. In previous work, the collaborative regulation of CO2 transport and fixation was investigated with co-overexpressing exogenous genes regulating both CO2 transport (sbtA and bicA) and PEP carboxylation (phosphoenolpyruvate (PEP) carboxylase (ppc) and carboxykinase (pck)) under trc promoter in Escherichia coli for succinate biosynthesis. For balancing metabolic flux to maximize succinate titer, a combinatorial optimization strategy to fine-tuning CO2 transport and fixation process was implemented by promoter engineering in this study. Firstly, based on the energy matrix a synthetic promoter library containing 20 rationally designed promoters with strengths ranging from 0.8% to 100% compared with the widely used trc promoter was generated. Evaluations of rfp and cat reporter genes provided evidence that the synthetic promoters were stably and had certain applicability. Secondly, four designed promoters with different strengths were used for combinatorial assembly of single CO2 transport gene (sbtA or bicA) and single CO2 fixation gene (ppc or pck) expression. Three combinations, such as Tang1519 (P4 -bicA + pP19 -pck), Tang1522 (P4 -sbtA + P4 -ppc), Tang1523 (P4 -sbtA + P17 -ppc) with a more than 10% increase in succinate production were screened in bioreactor. Finally, based on the above results, co-expression of the four transport and fixation genes were further investigated. Co-expression of sbtA, bicA, and ppc with weak promoter P4 and pck with strong promoter P19 (AFP111/pT-P4 -bicA-P4 -sbtA + pACYC-P19 -pck-P4 -ppc) provided the best succinate production among all the combinations. The highest succinate production of 89.4 g/L was 37.5% higher than that obtained with empty vector control. This work significantly enhanced succinate production through combinatorial optimization of CO2 transport and fixation. The promoter engineering and combinatorial optimization strategies used herein represents a powerful approach to tailor-making metabolic pathways for the production of other industrially important chemicals. Biotechnol. Bioeng. 2016;113: 1531-1541. © 2016 Wiley Periodicals, Inc.

Prevalence of personality disorders using two diagnostic systems in psychiatric outpatients in Shanghai, China: a comparison of uni-axial and multi-axial formulation.

OBJECTIVE: To compare multi-axial (DSM-IV) with uni-axial diagnostic system (CCMD-3, Chinese Classification and Diagnostic Criteria of Mental Disorders) as diagnostic methods to determine the prevalence of personality disorders (PDs) in Chinese psychiatric outpatients. METHOD: 3,075 outpatients were randomly sampled from clinical settings in China. CCMD-3 PDs were evaluated as per routine psychiatric practice. DSM-IV PDs were assessed using both self-reported questionnaire and structured clinical interview. RESULTS: The prevalence estimate for any type of PD in the total sample is 31.93% as reflected in the DSM-IV. This figure is nearly 110 times as large as the prevalence estimate for the CCMD-3. Only 9 outpatients were diagnosed with PD based on the CCMD-3. Amongst the 10 forms of DSM-IV PDs, avoidant (8.1%), obsessive-compulsive (7.6%), paranoid (6.0%), and borderline (5.8%) PDs were the most prevalent subtypes. This study found that PDs are commonly associated with the following: (i) the younger aged; (ii) single marital status; (iii) those who were not raised by their parents; (iv) introverted personalities; (v) first-time seekers of psycho-counseling treatment; and (vi) patients with co-morbid mood or anxiety disorders. CONCLUSIONS: PDs are easily overlooked when the diagnosis is made based on the CCMD-3 uni-axial diagnostic system. However, it was found that personality pathology is common in the Chinese psychiatric community when using the DSM-IV classification system. Existing evidence suggest, at least indirectly, that there are important benefits of moving towards a multi-axial diagnostic approach in psychiatric practice.

Teaching Chinese psychiatrists to make reliable dissociative disorder diagnoses.

The aim of the study was to assess the outcome of an educational effort by two North American experts in dissociative disorders to teach Chinese psychiatrists to make reliable dissociative disorder diagnoses. In the final phase of the educational effort, 569 patients at Shanghai Mental Health Center completed the Chinese version of the Dissociative Experiences Scale (DES). Patients were then randomly selected in different proportions according to their DES scores: 96 selected patients were then assessed with the Dissociative Disorders Interview Schedule (DDIS) and clinical diagnostic interviews based on DSM-IV criteria. According to the clinical diagnostic interviews, 28 (4.9%) patients were diagnosed as having dissociative disorders. Agreement between the American experts and Chinese psychiatrists for presence or absence of a dissociative disorder was 0.75 using Cohen's kappa. Dissociative disorders can be diagnosed in China with good inter-rater reliability. The authors describe the steps taken to achieve this outcome.

[A cross-sectional survey on personality disorder in mental disorder outpatients in Shanghai].

OBJECTIVE: To study the prevalence and risk factors for personality disorder (PD) outpatients attending in for psychiatric and psychological counseling in Shanghai. METHODS: 3075 subjects were sampled by systematic sampling method from outpatients in psycho-counseling clinics and psychiatric clinics in Shanghai Mental Health Center. Based on DSM-IV criteria, personality disorders were assessed by both questionnaires (personality diagnostic questionnaire, PDQ-4+) and interviews (structured clinical interview for DSM-IV Axis II, SCID-II). Logistic regression analysis was performed to determine the significant independent contributor to PD. RESULTS: 71.3% of the outpatients were found having pathological personality by using questionnaire of self rating PD scale. 982 outpatients (31.9%) met criteria for at least one personality disorder by using structured clinical interview. Younger age (OR = 1.8, 95%CI: 1.5 - 2.1), single or divorced (OR = 1.6, 95%CI: 1.4 - 1.9), psychological counseling outpatients (OR = 1.2, 95%CI: 1.1 - 1.3), mood and outpatients with neurosis disorders (OR = 1.7, 95%CI: 1.4 - 2.0) were more frequently assigned as personality disorders. Data from logistic regression analysis showed that patients of tender age, not nurtured and raised by their parents, with introvert characters were related risk factors of PD. CONCLUSION: High prevalence rate of PD was found in this sample of Chinese outpatients, especially in those psychological counseling outpatients with mood or neurosis disorders. More attention should be paid to the recognition and intervention of PD in outpatients with mental disorders.

Dissociative disorders among Chinese inpatients diagnosed with schizophrenia.

The purpose of this study was to assess the prevalence of dissociative disorders in a sample of Chinese psychiatric inpatients. Participants in the study were 569 consecutively admitted inpatients at Shanghai Mental Health Center, China, of whom 84.9% had a clinical diagnosis of schizophrenia based on the Chinese Classification and Diagnostic Criteria of Mental Disorders, Version 3. All participants completed a self-report measure of dissociation (the Dissociative Experiences Scale), and none had a prior diagnosis of a dissociative disorder. A total of 96 randomly selected participants were interviewed with a structured interview (the Dissociative Disorders Interview Schedule) and a clinical interview. These 96 patients did not differ significantly from the 473 patients who were not interviewed on any demographic measures or who did not complete the self-report dissociation measure. A total of 28 patients (15.3%, after weighting of the data) received a clinical diagnosis of a dissociative disorder based on Diagnostic and Statistical Manual of Mental Disorders (4th ed., text rev.) criteria. Dissociative identity disorder was diagnosed in 2 patients (0.53%, after weighting). Compared to the patients without a dissociative disorder, patients with dissociative disorders were significantly more likely to report childhood abuse (57.1% vs. 22.1%), but the 2 groups did not differ significantly on any demographic measures. Dissociative disorders were readily identified in an inpatient psychiatric population in China.