ObjectiveTo investigate the efficacy and safety of Kaixuan Jiedu compatibility and the decomposed prescriptions in the treatment of psoriasis. MethodsThirty Balb/c mice were randomly grouped as follows （n=6）： normal， model， Kaixuan Jiedu （KXJD， 15.21 g·kg^-1）， Kaixuan （KX， 3.08 g·kg^-1）， and Jiedu （JD， 12.13 g·kg^-1）. Except the normal group， the rest groups were modeled for psoriasis-like skin lesions by topical application of imiquimod， and samples were collected after 7 days of continuous intervention. Mice were photographed at the lesion site during modeling and before sampling and the psoriasis area and severity index （PASI） was calculated. Hematoxylin-eosin （HE） staining was used to observe pathological changes in the lesions and measure the epidermal thickness. Mice were photographed and observed for the tortuous dilation of dermal capillaries. The expression of vascular endothelial growth factor （VEGF）， platelet-endothelial cell adhesion molecule （CD31）， proliferating cell nuclear antigen （Ki67）， and cytokeratin 10 （CK10） in the epidermal tissue was detected by immunohistochemistry. Immunofluorescence assay was employed to determine the expression of Claudin-1 and Occludin. Real-time PCR was employed to determine the mRNA levels of interleukin-17A （IL-17A） and interleukin-23 （IL-23）. The spleen and thymus were photographed and weighed， and the spleen and thymus indices were calculated. The safety of the treatment was assessed by automatic biochemistry testing of the serum， liver， and kidney functions and by HE staining of the liver， kidney and spleen. ResultsCompared with that of the normal group， the skin of the model group showed erythema， infiltration， and typical psoriasis-like changes， tortuous dilation of dermal capillaries， hyperkeratosis in epidermal cells， acanthosis， massive lymphocytic infiltration in the dermis， impaired barrier function， increased expression of VEGF， CD31， Ki67， and CK10 （P<0.01）， reduced expression of Claudin-1 and Occludin （P<0.01） in the epidermis， and up-regulated mRNA levels of IL-17A and IL-23 （P<0.01）. In addition， the mice in the model group showed spleen enlargement， thymus atrophy， increased spleen index， and decreased thymus index （P<0.01）. Compared with the model group， KXJD and JD reduced psoriasis-like skin lesions， inhibited the tortuous dilation of dermal capillaries， reduced the expression of VEGF， CD31， Ki67， and CK10 （P<0.01）， increased the expression of claudin-1 （P<0.01）， and down-regulated the mRNA levels of inflammatory factors （P<0.01）. Moreover， the KXJD group outperformed the JD group. The JD group showed no significant difference from the model group regarding the spleen index， thymus index， and Occludin expression. The psoriasis indicators in the KX group were not significantly different from those in the model group. ConclusionKXJD and JD can reduce the symptoms of local skin lesions of psoriasis， which is manifested as different inhibition degrees of the proliferation and differentiation of keratin-forming cells， tortuous dilation of dermal capillaries， and inflammatory reactions， as well as the protection of the skin barrier. Moreover， KXJD outperformed JD. KX alone did not significantly reduce psoriasis lesions in mice. KXJD and the decomposed prescriptions are safe and effective， causing no obvious liver and kidney injuries.

ObjectiveTo investigate the efficacy and safety of Kaixuan Jiedu compatibility and the decomposed prescriptions in the treatment of psoriasis. MethodsThirty Balb/c mice were randomly grouped as follows （n=6）： normal， model， Kaixuan Jiedu （KXJD， 15.21 g·kg^-1）， Kaixuan （KX， 3.08 g·kg^-1）， and Jiedu （JD， 12.13 g·kg^-1）. Except the normal group， the rest groups were modeled for psoriasis-like skin lesions by topical application of imiquimod， and samples were collected after 7 days of continuous intervention. Mice were photographed at the lesion site during modeling and before sampling and the psoriasis area and severity index （PASI） was calculated. Hematoxylin-eosin （HE） staining was used to observe pathological changes in the lesions and measure the epidermal thickness. Mice were photographed and observed for the tortuous dilation of dermal capillaries. The expression of vascular endothelial growth factor （VEGF）， platelet-endothelial cell adhesion molecule （CD31）， proliferating cell nuclear antigen （Ki67）， and cytokeratin 10 （CK10） in the epidermal tissue was detected by immunohistochemistry. Immunofluorescence assay was employed to determine the expression of Claudin-1 and Occludin. Real-time PCR was employed to determine the mRNA levels of interleukin-17A （IL-17A） and interleukin-23 （IL-23）. The spleen and thymus were photographed and weighed， and the spleen and thymus indices were calculated. The safety of the treatment was assessed by automatic biochemistry testing of the serum， liver， and kidney functions and by HE staining of the liver， kidney and spleen. ResultsCompared with that of the normal group， the skin of the model group showed erythema， infiltration， and typical psoriasis-like changes， tortuous dilation of dermal capillaries， hyperkeratosis in epidermal cells， acanthosis， massive lymphocytic infiltration in the dermis， impaired barrier function， increased expression of VEGF， CD31， Ki67， and CK10 （P<0.01）， reduced expression of Claudin-1 and Occludin （P<0.01） in the epidermis， and up-regulated mRNA levels of IL-17A and IL-23 （P<0.01）. In addition， the mice in the model group showed spleen enlargement， thymus atrophy， increased spleen index， and decreased thymus index （P<0.01）. Compared with the model group， KXJD and JD reduced psoriasis-like skin lesions， inhibited the tortuous dilation of dermal capillaries， reduced the expression of VEGF， CD31， Ki67， and CK10 （P<0.01）， increased the expression of claudin-1 （P<0.01）， and down-regulated the mRNA levels of inflammatory factors （P<0.01）. Moreover， the KXJD group outperformed the JD group. The JD group showed no significant difference from the model group regarding the spleen index， thymus index， and Occludin expression. The psoriasis indicators in the KX group were not significantly different from those in the model group. ConclusionKXJD and JD can reduce the symptoms of local skin lesions of psoriasis， which is manifested as different inhibition degrees of the proliferation and differentiation of keratin-forming cells， tortuous dilation of dermal capillaries， and inflammatory reactions， as well as the protection of the skin barrier. Moreover， KXJD outperformed JD. KX alone did not significantly reduce psoriasis lesions in mice. KXJD and the decomposed prescriptions are safe and effective， causing no obvious liver and kidney injuries.

Background Lifestyle intervention or dietary modification has been the cornerstone of primary prevention and management of type 2 diabetes (T2D). Objective To investigate the associations of plant foods intake with the risk of incident T2D. Methods Based on a general population cohort, the Shanghai Suburban Adult Cohort and Biobank (SSACB), dietary data were collected for each participant in Songjiang District of Shanghai at enrollment with a validated Food Frequency Questionnaire (FFQ), and plant-based diet index (PDI), healthful plant-based diet index (hPDI), and unhealthful plant-based diet index (uPDI) were calculated. Incident T2D cases were identified according to physician diagnosis (fasting blood glucose ≥7.0 mmol·L⁻¹, or 2 h value during a 75-g oral glucose tolerance test ≥11.1 mmol·L⁻¹, or glycosylated hemoglobin ≥6.5%, or with typical symptoms of hyperglycemia or hyperglycemic crisis, accompanied by a random plasma glucose ≥11.1 mmol·L⁻¹) or medication records, obtained from the electronic information system for residents' medical insurance. Multivariable-adjusted Cox proportional hazards models and restricted cubic splines were used to evaluate the associations of foods from different sources with the risk of incident T2D. Results A total of 29016 participants [age at baseline (55.3±11.6) years] with a median follow-up duration of 5.688 years until 21 September 2022 were included. Plant foods (unprocessed) intake was associated with a decreased risk of incident T2D [HR (95%CI): 0.983 (0.969, 0.998)]. In comparison with participants in the highest quartile (≥859.3 g) of plant foods daily intake, the risk of incident T2D for those in the lowest quartile (<500.9 g) was higher [HR (95%CI): 1.250 (1.012, 1.544)]. No significant associations of animal foods [HR (95%CI): 1.006 (0.987, 1.026)] and processed foods [HR (95%CI): 0.978 (0.944, 1.014)] intakes were found with the risk of incident T2D. Replacing 50 g animal foods [HR (95%CI): 0.982 (0.968, 0.996)] or processed foods [HR (95%CI): 0.983 (0.969, 0.998)] with 50 g plant foods was associated with significantly decreased risks of incident T2D. Additionally, non-linear associations of PDI (P_nonlinear=0.023) and hPDI (P_nonlinear=0.016) with the risk of incident T2D were found in the SSACB. Conclusion Plant foods intake, especially healthful plant foods intake, is significantly associated with a decreased risk of incident T2D.

ObjectiveTo establish and validate a clinical prediction model for 1-year major adverse cardiovascular events（MACEs）risk after percutaneous coronary intervention （PCI） in coronary heart disease （CHD） patients with blood stasis syndrome. MethodThe consecutive CHD patients diagnosed with blood stasis syndrome in the Department of Integrative Cardiology at China-Japan Friendship Hospital from September 1， 2019 to March 31， 2021 were selected for a retrospective study， and basic clinical features and relevant indicators were collected. Eligible patients were classified into a derivation set and a validation set at a ratio of 7∶3， and each set was further divided into a MACEs group and a non-MACEs group. The factors affecting the outcomes were screened out by least absolute shrinkage and selection operator （Lasso） and used to establish a logistic regression model and identify independent prediction variables. The goodness-of-fit of the model was evaluated by the Hosmer-Lemeshow test， and the area under curve （AUC） of the receiver operating characteristic （ROC） curve， calibration curve， decision curve analysis （DCA）， and clinical impact curve （CIC） were employed to evaluate the discrimination， calibration， and clinical impact of the model. ResultA total of 731 consecutive patients were assessed and 404 eligible patients were enrolled， including 283 patients in the derivation set and 121 patients in the validation set. Lasso identified ten variables influencing outcomes， which included age， sex， fasting plasma glucose （FPG）， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C）， homocysteine （Hcy）， brachial-ankle pulse wave velocity （baPWV）， flow-mediated dilatation （FMD）， left ventricular ejection fraction （LVEF）， and Gensini score. The multivariate Logistic regression preliminarily identified age， FPG， TG， Hcy， LDL-C， LVEF， and Gensini score as the independent variables that influenced the outcomes. Of these variables， male， high FMD and high LVEF were protective factors， and the rest were risk factors. The prediction model for 1-year MACEs risk after PCI in CHD patients with blood stasis syndrome showed χ²=12.371 （P=0.14） in Hosmer-Lemeshow test and the AUC of 0.90. With the threshold probability > 10%， the model showed better prediction performance for 1-year MACEs risk after PCI in CHD patients with blood stasis syndrome than for that in all the patients. With the threshold probability > 60%， the estimated value was much closer to the real number of patients. ConclusionThe established clinical prediction model facilitates the early prediction of 1-year MACEs risk after PCI in CHD patients with blood stasis syndrome， which can provide ideas for the precise treatment of CHD patients after PCI and has guiding significance for improving the prognosis of the patients. Meanwhile， multi-center studies with larger sample sizes are expected to further validate， improve， and update the model.

OBJECTIVE: To carry out genetic testing for a patient with 45,X/46,XY mosaicism and autism spectrum disorder (ASD). METHODS: Peripheral blood samples of the patient and his parents were collected for the extraction of genomic DNA. Trio-based whole exome sequencing and Sanger sequencing were carried out thereafter. RESULTS: The proband and his father were found to harbor a heterozygous c.4781G>A (p.Arg1594Gln) variant of the CACNA1I gene. In addition, the proband was also found to harbor a de novo c.268C>T (p.Arg90Trp) missense variant of the MTRR gene. Based on guidelines of the American College of Medical Genetics and Genomics (ACMG), the c.4781G>A (p.Arg1594Gln) variant of the CACNA1I gene was predicted to be pathogenic (PVS1, PM1, PM2, PP3), while the c.268C>T (p.Arg90Trp) variant of the MTRR gene was predicted to be of uncertain significance. CONCLUSION Variants of the CACNA1I and MTRR genes, together with the chromosomal mosaicism, may have predisposed to the susceptibility to the ASD in this patient.
Autism Spectrum Disorder/genetics* ; Genomics ; Heterozygote ; Humans ; Mosaicism ; Whole Exome Sequencing

OBJECTIVE: To analyze the pathogenic variants of the KIF1A gene and its corresponding protein structure in an autism spectrum disorder (ASD) family trio carrying harmful missense variants in the KIF1A gene. METHODS: The peripheral blood DNA of the patient and his parents was extracted and sequenced using whole exome sequencing (WES) technology and verified by Sanger sequencing. Bioinformatics software SIFT, PolyPhen-2, Mutation Taster, and CADD software were used to analyze the harmfulness and conservation of variants. The Human Brain Transcriptome (HBT) database was used to analyze the expression of the KIF1A gene in the brain. PredictProtein and SWISS-MODEL were further used to predict the secondary structure and tertiary structure of KIF1A wild-type protein and variant protein. PyMOL V2.4 was utilized to investigate the change of hydrogen bond connection after protein variant. RESULTS: The WES sequencing revealed a missense variant c.664A>C (p.Asn222His) in the child's KIF1A gene, and this variant was a de novo variant. The harmfulness prediction results suggest that this variant is harmful. By analyzing expression level of KIF1A gene in the brain. It is found that KIF1A gene widely expressed in various brain regions during embryonic development. By analyzing the variant protein structure, the missense variant of KIF1A will cause many changes in the secondary structure of protein, such as alpha-helix, beta-strand, and protein binding domain. The connection of hydrogen bond and spatial structure will also change, thereby changing the original biological function. CONCLUSION The KIF1A gene may be a risk gene for ASD.
Autism Spectrum Disorder/genetics* ; Child ; Female ; Humans ; Kinesin/genetics* ; Mutation ; Mutation, Missense ; Pregnancy ; Protein Domains ; Whole Exome Sequencing

Diphtheria toxin is an ADP-ribosyltransferase toxic to human cells. Mutation of the active site in its catalytic domain eliminates the toxicity, but retains its immunogenicity. A non-toxic mutant of diphtheria toxin known as CRM197 protein has become an ideal carrier protein for conjugate vaccines. CRM197 can further improve its immunogenicity by cross-linking with other antigens, so it has good potential to find broad applications. Unfortunately, inclusion bodies are easily formed during the expression of recombinant CRM197 protein in Escherichia coli, which greatly reduces its yield. In order to address this problem, pG-KJE8 vector carrying molecular chaperones and plasmid pET28a-CRM197, were co-expressed in Escherichia coli. The results showed that the recombinant CRM197 protein was successfully expressed and appeared largely in inclusion bodies. The molecular chaperones DnaK, DnaJ, GrpE, GroES and GroEL5 expressed can facilitate correct and rapid folding of CRM197. Furthermore, it can also improve the recovery rate of soluble CRM197 protein. The soluble expression of CRM197 was maximized upon addition of 1.0 mmol/L IPTG, 0.5 mg L-arabinose, 5.0 ng/mL tetracycline and induction at 20oC for 16 h. The soluble CRM197 protein shows good immunoreactivity, demonstrating the molecular chaperones expressed from pG-KJE8 facilitated the soluble expression of CRM197 protein in E. coli.
Bacterial Proteins ; Diphtheria Toxin/genetics* ; Escherichia coli/genetics* ; Humans ; Molecular Chaperones/genetics* ; Recombinant Proteins/genetics*

Objective:To evaluate the effect of lycopene preconditioning on Toll-like receptors (TLRs)/nuclear factor kappa B (NF-κB) signaling pathway during hypoxia-reoxygenation (H/R) injury to rat cardiomyocytes.Methods:The rat H9C2 cardiomyocytes were cultured in vitro, inoculated in a petri dish at a density of 8×10 4 cells/ml and divided into 3 groups( n=30 each) using a random number table method: control group (C group), H/R group and lycopene preconditioning group (LP group). The H9C2 cardiomyocytes were reoxygenated for 6 h after hypoxia to establish a model of H/R injury in H/R group and LP group.The cells were preconditioned with lycopene 20 μmol/L at 12 h before establishing the model in LP group.The cell viability was detected by CCK-8 assay.The cell apoptosis rate was detected by flow cytometry (Annexin V and PI double staining). The levels of lactic dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyde (MDA) and reactive oxygen species (ROS) were detected by microplate method.The expression of TLR2, TLR3 and NF-κB was detected by Western blot. Results:Compared with group C, the cell viability and SOD level were significantly decreased, the apoptosis rate and levels of LDH, MDA and ROS were increased, and the expression of TLR2, TLR3 and NF-κB was up-regulated in H/R and LP groups ( P<0.05). Compared with group H/R, the cell viability and SOD level were significantly increased, the apoptosis rate and levels of LDH, MDA and ROS were decreased, and the expression of TLR2, TLR3 and NF-κB was down-regulated in group LP ( P<0.05). Conclusion:The mechanism by which lycopene preconditioning attenuates H/R injury may be related to inhibiting activation of TLRs/NF-κB signaling pathway and inhibiting oxidative stress response in rat cardiomyocytes.

Objective: To explore susceptibility genes for autism spectrum disorders (ASD). Methods: Whole-exome sequencing was carried out for 60 family trios affected with sporadic ASD. Genetic variants discovered in over 10% of the patients were selected for genotype-phenotype correlation and pathway enrichment analysis using Phenolyzer software and metascape database. Combining gene-phenotypic scores, pathway-related genes associated with neural and neurite triggering were screened for the candidates. Results: A total of 170 common variants were found to be associated with the ASD phenotype. Among these, there was only one high-confidence gene [SHANK2 (0.8146)] and four medium-confidence genes [ERBB2 (0.1322), LAMC3 (0.1117), PPFIA4 (0.1059), DISC1 (0.1002)]. Twenty-pathways and four biological processes were found to be statistically significant by pathway enrichment analysis, which included neuron projection morphogenesis (GO: 0048812), regulation of neuroblast proliferation (GO: 1902692), modulation of excitatory postsynaptic potential (GO: 0098815), and dendrite morphogenesis (GO: 0048813). Twenty-one genes were found to be closely associated with neurological and neurite triggering, among which only SHANK2, ERBB2, and DISC1 had above-medium confidence correlation scores with the ASD phenotypes. Conclusion Abnormal neuron projection morphogenesis (GO: 0048812) may be closely related to the occurrence of ASD. SHANK2, ERBB2, and DISC1 are susceptibility genes for ASD.

Objective To investigate the expression of connective tissue growth factor (CTGF) and heat shock protein 47 (HSP47) in peritoneum fibrosis rats,and the mechanism of 1,25-dihydroxyvitamin D3 [1,25-(OH)2-VitD3] in inhibiting the peritoneum fibrosis.Methods Adult male Sprague-Dawley rats were randomly divided into 3 groups:control group (n=8),model group (n=8) and 1,25-dihydroxyvitamin D3 group (VitD3,n=8).The model of peritoneum fibrosis rats were induced by daily intraperitoneally injection of 15％ chlorhexidine gluconate (CHX) 0.2 ml/d with 0.1％ glucose for 4 weeks.Rats in VitD3 group were also treated with 1,25-(OH)2-VitD3 [i.p.6 ng· (100 g) 1 · d-1].Peritoneal transport function,renal function,peritoneum thickness and serum level of 25hydroxyvitamin D3 were detected.In vitro,primary cultured peritoneal mesothelial cells were divided into control group,high glucose group (HG,2.5％),CTGF siRNA intervention group (CTGF siRNA+HG),VitD3 intervention group (VitD3+HG) and combined intervention group (CTGF siRNA+VitD3+HG).Real-time PCR,Western blotting and immunofluorescence were applied to measure the expression of CTGF and HSP47,also ELISA was used to detect the protein level of FN in peritoneum and peritoneal mesothelial cells.Results Compared with control group,the peritoneal ultrafiltration in peritoneum fibrosis rats were significantly decreased (P ＜ 0.05),the absorbance level of peritoneal fibrosis,peritoneum thickness,the rate of dialysate urea nitrogen and blood urea nitrogen (DUN/BUN) and the expressions of CTGF and HSP47 were increased (all P＜0.05).After application of 1,25-dihydroxyvitamin D3,peritoneal fibrosis lesion was significantly improved,the peritoneum thickness,the expressions of CTGF and HSP47 were decreased (all P ＜ 0.05).In vitro,2.5％ high glucose induced-peritoneal mesothelial cells were respectively treated by CTGF siRNA,1,25-(OH)2-VitD3 and combined interventions,the expression of FN,CTGF and HSP47 was significantly lower than that in high glucose group (all P ＜ 0.05).Conclusions The expression of CTGF and HSP47 is significantly increased in peritoneal fibrosis rats.1,25-(OH)2-VitD3 may ameliorate the progression of peritoneal fibrosis via reducing the expression of CTGF,decreasing the expression of HSP47 and FN.