ObjectiveMicrotube associated protein-2 （MAP-2）， alpha-tubulin （α-tubulin）， and synaptophysin （SYP） are important proteins in neuronal signal communication. This paper observed the effects of modified Zhigancao Tang on the expression of serum α-Synuclein （α-Syn） and its oligomers， MAP-2， α-tubulin， and SYP of patients with liver and kidney deficiency of Parkinson's disease （PD）， analyzed their correlation， and evaluated the therapeutic effect of modified Zhigancao Tang in patients with liver and kidney deficiency of PD based on α-Syn transmission pathway mediated by neuronal communication in vivo. MethodsA total of 60 patients with PD who met the inclusion criteria were randomly divided into a treatment group （30 cases） and a control group （30 cases）. Both groups were treated on the basis of PD medicine， and the treatment group was treated with modified Zhigancao Tang. Both groups were treated for 12 weeks. The changes in UPDRS score， TCM syndrome score， and expression of serum α-Syn and its oligomers， MAP-2， α-tubulin， and SYP were observed before and after 12 weeks of treatment in each group. The correlation between the above-mentioned serum biological indexes and the levels of serum α-Syn and its oligomers was analyzed. ResultsAfter treatment， the TCM syndrome score， UPDRS score， UPDRS-Ⅱ score， and UPDRS-Ⅲ score of the treatment group were significantly decreased （P<0.05， P<0.01）. The UPDRS score， UPDRS-Ⅱ score， and UPDRS-Ⅲ scores in the treatment group were significantly decreased compared with those in the control group after treatment （P<0.05）. After treatment， the total effective rate of the control group was 63.3% （19/30）， and that of the treatment group was 86.7% （26/30）. The clinical effect of the observation group was better than the control group （Z=-2.03， P<0.05）. The total effective rate of the observation group was better than that of the control group， and the difference was statistically significant （χ²=5.136， P<0.05）. After treatment， the oligomer level of serum α-Syn and MAP-2 level in the treatment group were significantly decreased （P<0.05， P<0.01）. The levels of serum α-Syn and its oligomers， as well as α-tubulin in the treatment group， were significantly decreased compared with those in the control group after treatment （P<0.05， P<0.01）. Serum α-Syn was correlated with serum MAP-2 and α-Syn oligomer in patients with PD （P<0.05， P<0.01） but not correlated with serum SYP . Serum α-Syn oligomers of patients with PD were correlated with serum MAP-2 and α-tubulin （P<0.05， P<0.01） but not correlated with serum SYP level. Serum SYP of patients with PD was correlated with serum MAP-2 （P<0.05）. ConclusionModified Zhigancao Tang has a therapeutic effect on patients with liver and kidney deficiency of PD by inhibiting the production of α-Syn oligomers and intervening α-Syn microtubule transport pathway in vivo.

BACKGROUND: Islet transplantation is currently considered the most promising method for treating insulin-dependent diabetes. The two most-studied artificial islets are alginate-encapsulated b cells or b cell spheroids. As three-dimensional (3D) models, both artificial islets have better insulin secretory functions and transplantation efficiencies than cells in twodimensional (2D) monolayer culture. However, the effects of these two methods have not been compared yet. Therefore, in this study, cells from the mouse islet b cell line Min6 were constructed as scaffold-free spheroids or alginate-encapsulated dispersed cells. METHODS: MIN6 cell spheroids were prepared by using Agarose-base microwell arrays. The insulin secretion level was determined by mouse insulin ELISA kit, and the gene and protein expression status of the MIN6 were performed by Quantitative polymerase chain reaction and immunoblot, respectively. RESULTS: Both 3D cultures effectively promoted the proliferation and glucose-stimulated insulin release (GSIS) of MIN6 cells compared to 2D adherent cells. Furthermore, 1% alginate-encapsulated MIN6 cells demonstrated more significant effects than the spheroids. In general, three pancreatic genes were expressed at higher levels in response to the 3D culture than to the 2D culture, and pancreatic/duodenal homeobox-1 (PDX1) expression was higher in the cells encapsulated in 1% alginate than that in the spheroids. A western blot analysis showed that 1% alginate-encapsulated MIN6 cells activated the phosphoinositide 3-kinase (PI3K)/serine/threonine protein kinase (AKT)/forkhead transcription factor FKHR (FoxO1) pathway more than the spheroids, 0.5% alginate-, or 2% alginate-encapsulated cells did. The 3D MIN6 culture, therefore, showed improved effects compared to the 2D culture, and the 1% alginate-encapsulated MIN6 cells exhibited better effects than the spheroids. The upregulation of PDX1 expression through the activation of the PI3K/AKT/FoxO1 pathway may mediate the improved cell proliferation and GSIS in 1% alginate-encapsulated MIN6 cells. CONCLUSION This study may contribute to the construction of in vitro culture systems for pancreatic islets to meet clinical requirements.

Objective:To study the value of neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) in diagnosing severe Mycoplasma pneumoniae pneumonia (MPP).Methods:A total of 616 cases of MPP patients in the Children′s Hospital of Soochow University from January 2015 to December 2017 were retrospectively analyzed.During the same period, 100 healthy children were selected as the healthy control group.NLR and PLR between MPP group and healthy control group, and those between severe MPP group and ordinary MPP group were compared by t test or rank sum test.Risk factors for severe MPP were identified.Receiver operating characteristic(ROC) curves were plotted to identify the cut-off point of NLR and PLR in distinguishing MPP from healthy subjects. Results:(1)The median of white blood cell count (WBC), neutrophil count (N), platelet count (PLT), NLR, PLR, immunoglobulin M (IgM) and the median percentage of CD3 -CD 19+ , CD 19+ CD 23+ in MPP group were significantly higher than those in healthy control group(8.36×10 9/L vs.7.49×10 9/L, 4.41×10 9/L vs.3.11×10 9/L, 340.92×10 9/L vs.234.00×10 9/L, 1.70 vs.0.91, 112.99 vs.70.34, 1.33 g/L vs.1.29 g/L, 20.95% vs.17.10%, 11.25% vs.9.70%), whereas the median of lymphocyte count (L), IgA and the median percentage of CD3 + , CD3 + CD8 + , and CD3 -CD +(16+ 56) were significantly lower(2.64×10 9/L vs.3.37×10 9/L, 0.86 g/L vs.1.30 g/L, 64.55% vs.68.00%, 23.65% vs.24.90%, 10.50% vs.12.20%)( Z=-3.074, -2.413, -2.972, -1.357, -1.863, -2.251, -4.282, -3.420, -2.221, -4.181, -2.784, -2.024, -2.791, all P<0.05). (2)The median of N, NLR, PLR, IgA, IgG, IgM and the average of percentage of CD3 + , CD3 + CD8 + in severe MPP group were significantly higher than those in ordinary MPP group[5.18×10 9/L vs.3.52×10 9/L, 2.39 vs.1.03, 149.32 vs.94.23, 1.29 g/L vs.0.71 g/L, 9.63 g/L vs.8.19 g/L, 1.40 g/L vs.1.29 g/L, (65.53±9.75)% vs.(62.81±9.89)%, (25.35±6.65)% vs.(23.38±6.91)%], whereas the median of L, the median percentage of CD3 -CD 19+ , and CD 19+ CD 23+ were significantly lower than those of ordinary MPP group(2.02×10 9/L vs.3.25×10 9/L, 17.40% vs.21.50%, 9.00% vs.11.70%)( Z/ t=-7.807, -11.313, -10.452, -8.819, -6.162, -3.047, -3.128, -3.270, -9.402, -5.191, -5.214, all P<0.05). (3)Univariate and multivariate Logistic regression analysis showed that CD3 -CD 19+ was the protective factor for severe MPP, while N, NLR and PLR were the risk factors for severe MPP (all P<0.05), with the risk sequence of NLR>PLR>N.(4)Area under ROC curve analysis of NLR and PLR in the diagnosis of severe MPP: NLR: AUC=0.789, 95% CI: 0.754~0.823, P<0.001; PLR: AUC=0.767, 95% CI: 0.730~0.804, P<0.001; when the critical value of NLR was 1.09, the sensitivity was 98.9%, and the specificity was 70.6%.When the critical value of PLR was 97.47, the sensitivity and specificity were 88.5% and 69.4%. Conclusions:NLR and PLR can be served as independent influencing factors for severe MPP, showing the diagnostic potential in severe MPP.

Objective:To clarify the function and molecular mechanisms of serpin family E member 2 (SERPINE2) in cellular migration and invasion of esophageal squamous cell carcinoma (ESCC).Methods:The expression of SERPINE2 in ESCC was analyzed by using online databases TCGA (http: //gepia.cancer-pku.cn/detail.php and http: //ualcan.path.uab. edu/index.html). The expressions of SERPINE2 mRNA in normal human esophageal epithelial cell line NE2, human ESCC cell lines KYSE30 and KYSE150 were detected by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). SERPINE2-konckdown or SERPINE2-overexpressed plasmid was transfected into KYSE30 cells, and the efficiencies of the knockdown and overexpression system were tested by qRT-PCR. The relationships of SERPINE2 and ESCC migration and invasion were determined by migration and invasion assays in vitro. The associations between SERPINE2 expression and β-catenin as well as its target genes including c-Myc, cyclin D1 and CD44 were analyzed by immunofluorescence, qRT-PCR and western blot, respectively.Results:The expressions of SERPINE2 were significantly upregulated in both esophageal cancer (ESCA) and ESCC tissues compared to normal tissues by analyzing 182 and 95 cases, respectively ( P<0.01). SERPINE2 is highly expressed in both KYSE30 and KYSE150 cells ( P<0.05). The number of migrating and invading cells in control group were (212.66±24.11)/field and (136.00±14.42)/field, while were (88.33±9.71)/field and (77.00±9.53)/field in SERPINE2-knockdown 1 group, and (66.00±8.00)/field and (45.66±3.78)/field in SERPINE2-knockdown 2 group, respectively, and the differences were dramatically significant compared with the control group ( P<0.01). The number of migrating and invading cells in control group were (250.00±30.00)/field and (203.33±15.27)/field, while were (383.33±35.11)/field and (246.66±25.16)/field in SERPINE2-overpressed group, and the differences were strikingly significant compared with the control group ( P<0.01). The protein expression of β-catenin was upregulated while phosphorylated β-catenin protein expression was downregulated in SERPINE2-overexpressed KYSE30 cells when compared to control cells.The transcription activity of β-catenin was significantly upregulated and the mRNA expressions of its target genes including c-Myc, cyclin D1 and CD44 were all increased. After treated with 25 μM iCRT14, the number of migrated cells in the control and SERPINE2-overpressed groups were (200.00±36.05)/field and (258.33±22.54)/field, and the number of invaded cells were (160.00±17.32)/field and (188.33±25.65)/field, respectively, the differences were dramatically significant compared with the group without iCRT14 treatment ( P<0.01). Conclusion:SERPINE2 is significantly upregulated in ESCC cells and can promote cellular migration and invasion by activating β-catenin, which may provide a potential therapeutic target for patients with ESCC.

The development of new biomarkers or therapeutic targets for cancer immunotherapies requires deep under-standing of T cells. To date, the complete landscape and systematic characterization of long noncoding RNAs (lncRNAs) in T cells in cancer immunity are lacking. Here, by systematically analyzing full-length single-cell RNA sequencing (scRNA-seq) data of more than 20,000 libraries of T cells across three cancer types, we provided the first comprehensive catalog and the functional repertoires of lncRNAs in human T cells. Specifically, we developed a custom pipeline for de novo transcriptome assembly and obtained a novel lncRNA catalog containing 9433 genes. This increased the number of current human lncRNA catalog by 16％and nearly doubled the number of lncRNAs expressed in T cells. We found that a portion of expressed genes in single T cells were lncRNAs which had been overlooked by the majority of previous studies. Based on metacell maps constructed by the MetaCell algorithm that partitions scRNA-seq datasets into disjointed and homogenous groups of cells (metacells), 154 signature lncRNA genes were identified. They were associated with effector, exhausted, and regulatory T cell states. Moreover, 84 of them were functionally annotated based on the co-expression networks, indicating that lncRNAs might broadly participate in the regulation of T cell functions. Our findings provide a new point of view and resource for investigating the mechanisms of T cell regulation in cancer immunity as well as for novel cancer-immune biomarker development and cancer immunotherapies.

Objective:To clarify the function and molecular mechanisms of serpin family E member 2 (SERPINE2) in cellular migration and invasion of esophageal squamous cell carcinoma (ESCC).Methods:The expression of SERPINE2 in ESCC was analyzed by using online databases TCGA (http: //gepia.cancer-pku.cn/detail.php and http: //ualcan.path.uab. edu/index.html). The expressions of SERPINE2 mRNA in normal human esophageal epithelial cell line NE2, human ESCC cell lines KYSE30 and KYSE150 were detected by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). SERPINE2-konckdown or SERPINE2-overexpressed plasmid was transfected into KYSE30 cells, and the efficiencies of the knockdown and overexpression system were tested by qRT-PCR. The relationships of SERPINE2 and ESCC migration and invasion were determined by migration and invasion assays in vitro. The associations between SERPINE2 expression and β-catenin as well as its target genes including c-Myc, cyclin D1 and CD44 were analyzed by immunofluorescence, qRT-PCR and western blot, respectively.Results:The expressions of SERPINE2 were significantly upregulated in both esophageal cancer (ESCA) and ESCC tissues compared to normal tissues by analyzing 182 and 95 cases, respectively ( P<0.01). SERPINE2 is highly expressed in both KYSE30 and KYSE150 cells ( P<0.05). The number of migrating and invading cells in control group were (212.66±24.11)/field and (136.00±14.42)/field, while were (88.33±9.71)/field and (77.00±9.53)/field in SERPINE2-knockdown 1 group, and (66.00±8.00)/field and (45.66±3.78)/field in SERPINE2-knockdown 2 group, respectively, and the differences were dramatically significant compared with the control group ( P<0.01). The number of migrating and invading cells in control group were (250.00±30.00)/field and (203.33±15.27)/field, while were (383.33±35.11)/field and (246.66±25.16)/field in SERPINE2-overpressed group, and the differences were strikingly significant compared with the control group ( P<0.01). The protein expression of β-catenin was upregulated while phosphorylated β-catenin protein expression was downregulated in SERPINE2-overexpressed KYSE30 cells when compared to control cells.The transcription activity of β-catenin was significantly upregulated and the mRNA expressions of its target genes including c-Myc, cyclin D1 and CD44 were all increased. After treated with 25 μM iCRT14, the number of migrated cells in the control and SERPINE2-overpressed groups were (200.00±36.05)/field and (258.33±22.54)/field, and the number of invaded cells were (160.00±17.32)/field and (188.33±25.65)/field, respectively, the differences were dramatically significant compared with the group without iCRT14 treatment ( P<0.01). Conclusion:SERPINE2 is significantly upregulated in ESCC cells and can promote cellular migration and invasion by activating β-catenin, which may provide a potential therapeutic target for patients with ESCC.

To investigate the social support level and its influencial factors in patients with systemic lupus erythematosus (SLE), and to develop the management strategies for chronic disease.
 Methods: Patients with SLE were investigated by Social Support Rating Scale (SSRS), Patient Health Questionnaire (PHQ-9), Generalized Anxiety Disorder (GAD-7), 36-Item Short-Form Health Survey (SF-36) and Visual Analogue Scale (VAS) of fatigue. The demographic and clinical data of SLE patients were recorded. SLE disease activity and damage severity were assessed by SLE Disease Activity Index (SLEDAI) and SLE Damage Index (SDI), respectively. Influencial factors for social support were analyzed.
 Results: A total of 246 patients were included. Social support scores for these patients were 40.76±7.93 and the scores showed no significant difference with the national norm (P>0.05). Patients who were younger than 18, single, unemployed or damaged by disease showed lower level of social support (P<0.05). Compared with the high social support group, patients in the low social support group experienced more severe depression or anxiety, and scored lower on mental component summary scale (vitality, social functioning, emotional role and mental health perception) and physical role of SF-36 (P<0.05).
 Conclusion: Social support levels for patients with SLE are closely related to the quality of life, and influenced by age, marital status, professional condition, and disease damage. Health education for patients and their families should be strengthened in chronic disease management to enhance social support and finally, improve their quality of life.
Chronic Disease ; Health Status ; Humans ; Lupus Erythematosus, Systemic ; Quality of Life ; Severity of Illness Index ; Social Support ; Surveys and Questionnaires

Objective To investigate the inhibiting effect of CGP77675 (CGP),a Src inhibitor,on epithelial-mesenchymal transition (EMT) of human retinal pigment epithelial (RPE) cells induced by transformation growth factor-β1 (TGF-β1).Methods Human RPE cell line (ARPE19 cells) was cultured in vitro and divided into control group,TGF-β1 group and TGF-β1 +CGP group.Corresponding agent was added into culture medium based on grouping.The morphology of the cells were examined under the optical microscope 3 days after culture.The expressions of EMT-related genes and proteins in the cells were detected by real-time quantitative PCR and Western blot,respectively,including fibronectin 1 (FN 1),and plasminogen activation inhibitor 1 (PAI1),and the expressions of zonula occludens protein 1 (ZO1) and cytoskeleton protein filamentous actin (F-actin) were detected by immunofluorescence staining.MTT assay was employed to evaluate the cell proliferation rate.The migration distance of the cells was measured by scratch test.Results The ARPE19 cells in the control group showed an epithelial-like morphology and F-actin and ZO-1 were expressed along cell membrane.In the TGF-β1 group,the cells appeared to be fibrous-like,and the fluorescence staining of F-actin was disordered and ZO-1 was discontinuous on the cell membrane.The cells in the TGF-β1 +CGP group remained to be an epithelial-like in shape with clear and complete expressions of F-actin and ZO-1.The relative expressions of FN1 mRNA and PAI1 mRNA in the cells were 0.211 ± 0.080 and 0.116±0.073,1.000±0.001 and 1.000±0.001,0.368±0.097 and 0.362±0.048 in the control group,TGF-β1 group and TGF-β1 +CGP groups,showing significant differences among the groups (F=33.14,82.92;both at P＜0.01),with the highest expressions ofFN1 mRNA and PAI1 mRNA in the TGF-β1 group (all at P＜0.05).The relative expressions of FN1 and PAI1 proteins were 0.166±0.055 and 0.327±0.066,1.000±0.001 and 1.000± 0.001,0.143 ± 0.030 and 0.260 ± 0.077 in the control group,TGF-β1 group and TGF-β1 + CGP group,with significant differences among three groups (F=181.90,48.85;both at P＜0.01),and the expressions FN1 and PAI1 proteins were significantly higher in the TGF-β1 than those in the control group and TGF-β1 +CGP group (all at P＜0.05).The cell proliferative rate in the TGF-β1+CGP group was (79.30±3.44) ％ and (54.80±7.39) ％ at the third day and seventh day after culture,which were significantly reduced in comparison with (99.50 ± 1.00)％ and (99.10±0.50)％ in the control group as well as (95.10±4.20)％ and (92.10±4.50)％ in the TGF-β1 group (all at P＜0.05).The migration distance was disappeared in the TGF-β1 group,and the scratch width was not obviously changed in the TGF-β1 +CGP group.Conclusions Src inhibitor can inhibit EMT process of ARPE19 cells induced by TGF-β1,indicating that Src signaling pathway may play a critical role in EMT of RPE cells.

Objective To investigate the role of the ATP binding box C subfamily 2 transporter protein (ABCC2) gene in drug-resistant epilepsy.Methods From February 2014 to February 2018,204 epileptic patients were treated in our hospital,including 41 cases of drug resistance epilepsy (drug resistance group),163 cases of non drug resistant epilepsy (sensitive group).The rs717620 polymorphism of ABCC2 gene and the level of P-glycoprotein (P-gp) in cerebrospinal fluid and serum were detected in two groups.Results The proportion of genotype TT and gene frequency T in the drug resistant group were 24.39％ and 41.46％,respectively,which were significantly higher than those in the sensitive group (P ＜ 0.05).The concentration of P-gp in cerebrospinal fluid of drug resistant group was (21.03 ± 4.21) ng/ml,which was significantly higher than that of the sensitive group (P ＜ 0.05).There was no significant difference in serum P-gp concentration between the drug resistant group and the sensitive group (P ＞ 0.05);The concentration of P-gp in cerebrospinal fluid of patients with genotype TT in drug resistance group was (24.03 ± 3.57) ng/ml,which was significantly higher than that of type CC and CT (P ＜ 0.05).There was no significant difference in the concentration of P-gp in the patients with different genotypes in the drug resistant group (P ＞ 0.05).Conclusions The rs717620 polymorphism of ABCC2 gene may be associated with drug-resistant epilepsy,and may be related to P-gp level in the cerebrospinal fluid.

Objective To investigate the prevalence of sleep disorders and the relevant determinants in a cohort of systemic lupus erythematosus (SLE) patients.Methods One hundred patients with SLE were included in the study.Sleep quality was assessed using the Pittsburgh sleep quality index (PSQI).Depression,anxiety,quality of life,and fatigue were evaluated by patient health questionnaire (PHQ)-9,generalized anxiety disorder (GAD)-7,short form 36 health survey (SF-36),and visual analogue scale (VAS) respectively.The demographic and clinical data were also recorded.SLE disease activity and damage severity were assessed by systemic lupus erythematosus disease activity index (SLEDAI) and systemic lupus erythematosus damage index (SDI) respectively.Mann-Whitnry U test,t test,Logistic regression were used for statistically analysis.Results The prevalence of sleep disorders in SLE patients was 42％.Compared with patients without sleep disorders,the ratio of males and married patients,age,the score of SDI,PHQ-9,GAD-7,and fatigue were higher in SLE patients with sleep disorders,while the score of SF-36 was lower (r＜0.05).Age,SLEDAI,SDI,PHQ-9,GAD-7,and fatigue correlated positively with sleep disorders (The values of r were 0.215,0.230,0.311,0.529,0.455,0.541,P＜0.05).C3 and the score of SF-36 correlated negatively with sleep disorders (The values of r were-0.204,-0.342,-0.490,-0.464,-0.497,-0.590,-0.428,-0.478,-0.398,-0.412,-0.659respectively,P＜0.05).In multi-ple logistic regression analyses,gender (OR=22.22),anxiety (OR =2.895),body pain (OR =0.964),and energy (OR =0.947) were the independent determinants of sleep disorders (R2=0.494,P＜0.01).Conclusion Poor sleep quality is common in SLE patients.Gender,age,disease activity and severity,anxiety,depressed mood,and quality of life contribute significantly to sleep disorders in SLE.