Homo- or heterodimeric compounds that affect dimeric protein function through interaction between monomeric moieties and protein subunits can serve as valuable sources of potent and selective drug candidates. Here, we screened an in-house dimeric natural product collection, and panepocyclinol A (PecA) emerged as a selective and potent STAT3 inhibitor with profound anti-tumor efficacy. Through cross-linking C712/C718 residues in separate STAT3 monomers with two distinct Michael receptors, PecA inhibits STAT3 DNA binding affinity and transcription activity. Molecular dynamics simulation reveals the key conformation changes of STAT3 dimers upon the di-covalent binding with PecA that abolishes its DNA interactions. Furthermore, PecA exhibits high efficacy against anaplastic large T cell lymphoma in vitro and in vivo, especially those with constitutively activated STAT3 or STAT3^Y640F. In summary, our study describes a distinct and effective di-covalent modification for the dimeric compound PecA to disrupt STAT3 function.

Objective:To explore the prognostic value of BRCA1-associated protein 1 (BAP1) expression loss in patients with malignant mesothelioma (MM) .Methods:A total of 82 MM patients from January 1998 to December 2017 in Zhejiang Province were selected to detect the expression of BAP1 protein by immunohistochemical analysis. Kaplan-Meier method was used to draw the survival curve, and multivariate Cox proportional risk model was used to analyze the factors affecting the survival rate.Results:Among 82 MM patients, 61 (74.4%) were female, aged (57±11) years. BAP1 protein expression was deficient in 39 patients (47.6%). The survival rate was correlated with the loss of BAP1 protein expression and age (χ 2=5.27, 5.66, P=0.022, 0.017). Subgroup analysis showed that loss of BAP1 protein expression was associated with better prognosis in MM patients <57 years of age, female, pleural MM, epithelial MM, and treated with drugs or surgery ( P<0.05). Multivariate model results showed that positive expression of BAP1 protein ( HR=3.75, 95% CI: 2.23-6.30, P<0.001) and age ≥57 years ( HR=1.66, 95% CI: 1.01-2.72, P=0.049) were risk factors for survival in patients with MM. Conclusion:Loss of BAP1 protein expression may be an independent prognostic factor in patients with MM, which is associated with longer survival.

Objective:To explore effect of IgG receptor FcγRⅡB on neuronal injury and imbalance of Th17/Treg in experi-mental autoimmune encephalomyelitis(EAE)model mice.Methods:C57BL/6 mice were randomly divided into control group,EAE group,FcγRⅡB group and EAE+FcγRⅡB group,with 15 mice in each group.EAE model was induced by subcutaneous injection of MOG35-55 peptide and treated with FcγRⅡB lentiviral solution.After modeling was established,body weight of mice was weighed every day,and neurological function was scored for 30 d;after 30 days,mice were sacrificed.HE staining was used to observe patho-logical changes of brain tissue,LFB staining was used to assess structural changes of spinal cord myelin,and immunofluorescence staining was used to detect spinal cord cerebral cortex neuron nuclear antigen(NeuN)and Caspase-3 expressions,TUNEL staining was used to detect apoptosis of neurons,ELISA was used to detect serum IL-6,IL-17,IL-10 and TGF-β levels,flow cytometry was used to analyze proportion of Th17 and Treg cells in spleen,Western blot was used to determine protein expressions of retinoic acid-related orphan receptor γt(RORγt)and Forkhead family transcription factor 3(Foxp3)in spinal cord tissue.Results:Compared with control group,mice in EAE group had decreased body weight,increased neurological function scores,obvious infiltration of inflamma-tory cells in brain tissue,and signs of demyelination in spinal cord,fluorescence expression intensity of NeuN was weakened and fluorescence expression intensity of Caspase-3 was enhanced,there were more TUNEL-positive stained cells,number of apoptotic cells was increased,levels of IL-6 and IL-17 in serum were increased,and levels of IL-10 and TGF-β were decreased,proportion of Th17 cells in spleen was increased,proportion of Treg was decreased,expression of RORγt protein in spinal cord tissue was up-regu-lated while relative expression of Foxp3 protein was down-regulated(P<0.05);compared with EAE group,weight of mice in EAE+FcγRⅡB group was increased,neurological function score was decreased,infiltration of inflammatory cells in brain tissue was reduced,demyelination of spinal cord was improved,fluorescence expression intensity of NeuN was enhanced,and fluorescence expression intensity of Caspase-3 was weakened,there were fewer TUNEL-positive stained cells,number of apoptotic cells was decreased,levels of IL-6 and IL-17 in serum were decreased,while levels of IL-10 and TGF-β were increased,at the same time,proportion of Th17 cells in spleen was decreased and proportion of Treg was increased,expression of RORγt protein in spinal cord tissue was down-regulated,while expression of Foxp3 protein was up-regulated(P<0.05).Conclusion:FcγRⅡB has neuroprotective effect on EAE mice,and can reduce infiltration of inflammatory cells and demyelination in brain tissue,whose mechanism may be related to regulation of cytokine levels and immune balance of Th17/Treg cells.

Objective:To investigate the effect of glucocorticoids on brain injury in rats with Streptococcus pneumoniae(SP)meningitis,as well as the changes of cerebrospinal fluid immunoglobulin levels under this effect.Methods:A total of 40 SD rats were divided into control group,model group,low-dose methylprednisolone sodium succinate group(Low-MSS)and high-dose methylpred-nisolone sodium succinate group(High-MSS)according to random table method,with 10 rats in each group.Except for the control group,SP meningitis models were established in other groups.Rats in Low-MSS group and High-MSS group were injected with methyl-prednisolone sodium succinate by tail vein at doses of 5 mg/kg and 10 mg/kg,respectively,once a day for 21 days.Neurobehavioral scores of rats after modeling were performed by Loeffler method;contents of immunoglobulin IgA,IgM and IgG in cerebrospinal fluid of rats were detected by immunoturbidimetry;levels of inflammatory cytokines TNF-α,IL-6 and IFN-γ in supernatant of rats brain ho-mogenate were determined by ELISA;HE staining was used to detect pathological changes of rats brain tissue;Nissl staining was used to detect the number of neuronal cells in rats brain tissue;TUNEL staining was used to detect the number of apoptotic cells in rats brain tissue;immunofluorescence staining was used to detect expressions of GFAP and S-100b,the marker proteins of rat brain tissue injury.Results:Before SP injection,there was no difference in neurobehavioral scores between control group and model group(P>0.05),at 24 h,48 h and 72 h after SP injection,neurobehavioral scores in model group were significantly lower than those in control group(P<0.05).Compared with control group,contents of IgA,IgM and IgG in cerebrospinal fluid of rats in model group were in-creased,and contents of TNF-α,IL-6 and IFN-γ in supernatant of the brain homogenate were also increased,the tissue was obviously damaged,accompanied by a large number of inflammatory cells infiltration,the number of neuronal cells were decreased,the number of apoptotic cells were increased,and the fluorescence density values of S-100b and GFAP were increased(P<0.05);compared with model group,contents of IgA,IgM and IgG in cerebrospinal fluid of rats in Low-MSS group and High-MSS group were decreased,and contents of TNF-α,IL-6 and IFN-γ in brain homogenate supernatant were also decreased(P<0.05),brain tissue damage was alleviated,inflammatory cell infiltration was significantly reduced,neuronal cells were increased,the number of apoptotic cells were decreased,and the fluorescence density values of S-100b and GFAP were also decreased(P<0.05).In addition,the improvement effect of various indexes and brain tissue damage in High-MSS group was better than that in Low-MSS group,and the difference were statistically signifi-cant(P<0.05).Conclusion:Glucocorticoid methylprednisolone sodium succinate can effectively improve the brain tissue damage in SP meningitis rats,regulate the levels of immunoglobulin IgA,IgM and IgG,and has a protective effect on SP meningitis rats.

Objective:To explore the prognostic value of BRCA1-associated protein 1 (BAP1) expression loss in patients with malignant mesothelioma (MM) .Methods:A total of 82 MM patients from January 1998 to December 2017 in Zhejiang Province were selected to detect the expression of BAP1 protein by immunohistochemical analysis. Kaplan-Meier method was used to draw the survival curve, and multivariate Cox proportional risk model was used to analyze the factors affecting the survival rate.Results:Among 82 MM patients, 61 (74.4%) were female, aged (57±11) years. BAP1 protein expression was deficient in 39 patients (47.6%). The survival rate was correlated with the loss of BAP1 protein expression and age (χ 2=5.27, 5.66, P=0.022, 0.017). Subgroup analysis showed that loss of BAP1 protein expression was associated with better prognosis in MM patients <57 years of age, female, pleural MM, epithelial MM, and treated with drugs or surgery ( P<0.05). Multivariate model results showed that positive expression of BAP1 protein ( HR=3.75, 95% CI: 2.23-6.30, P<0.001) and age ≥57 years ( HR=1.66, 95% CI: 1.01-2.72, P=0.049) were risk factors for survival in patients with MM. Conclusion:Loss of BAP1 protein expression may be an independent prognostic factor in patients with MM, which is associated with longer survival.

Objective : To examine the diagnostic and prognostic value of long non-coding RNA (lncRNA) JPX in mesothelioma, so as to provide insights into diagnosis and prognosis of mesothelioma. Methods: Patients with clinically definitive diagnosis of mesothelioma from 2015 to 2019 that were sampled from asbestos processing plants in Zhejiang Province from 2015 to 2019 were recruited in the mesothelioma group, while healthy residents without asbestos exposure or asbestos-related diseases in the same area served as controls. Participants' demographics, pathologic diagnosis and imaging features were collected, and the expression of blood lncRNA JPX was detected using lncRNA microarrays. The diagnostic value of lncRNA JPX for mesothelioma was evaluated using the receiver operating characteristic (ROC) curve, and the correlation between lncRNA JPX expression and prognosis was examined among mesothelioma patients using survival analysis. Results: There were 17 subjects in the mesothelioma group, with a mean age of (65.71±8.36) years, and 34 subjects in the controls, with a mean age of (64.24±8.70) years. LncRNA microarray detected significantly high lncRNA JPX expression in mesothelioma patients, and higher blood lncRNA JPX expression was detected in the mesothelioma group than in the control group [median (interquartile range), 1.10 (1.31) vs. 0.89 (0.54); t'=-2.300, P=0.034]. The area under the ROC curve was 0.673 (95%CI: 0.507-0.839, P=0.046), and if the cutoff was 1.759, the sensitivity and specificity were 35.3% and 100.0%, respectively. Survival analysis showed no significant difference in the survival rate of mesothelioma patients between the high lncRNA JPX expression group and the low expression group (χ2=0.212, P=0.645). Conclusions LncRNA JPX overexpression is detected in the blood of patients with mesothelioma, and lncRNA JPX expression presents a diagnostic value for mesothelioma; however, it shows little prognostic value for mesothelioma.

Objective:To investigate the regulation of intestinal microbiota by Roux-en-Y gastric bypass (RYGB) on patients with obesity or obesity combined with diabetes.Methods:The retrospective and descriptive study was conducted. The stool samples before and after surgery and clinical data of 20 patients with obesity, including 9 simple obesity cases and 11 obesity combined with diabetes cases, who underwent RYGB in the First Affiliated Hospital of Ji′nan University from July 2016 to August 2017 were collected. There were 11 males and 9 females, aged (33±11)years. Observation indicators: (1) changes in composition and structure of intestinal microflora; (2) changes of intestinal microflora in simple obesity patients after operation; (3) changes of intestinal microflora in obesity combined with diabetes patients after operation. Follow up was conducted using telephone interview or outpatient examinations to detect the body mass, the application of antimicrobial agent and the blood glucose control of patients. According to the unified training points, the stool samples were collected and stored into the DNA stabilizer, and then conducted to laboratory analysis within 45 hours. The follow up was up to November 2018. Measurement data with normal distribution were represented as Mean± SD, and independent-samples t test was used for inter-group comparison and paired-samples t test was used for intra-group comparison. Measurement data with skewed distribution were represented as M( Q1, Q3), and Wilcoxon signed rank test of two independent samples was used for inter-group comparison. Count data were described as absolute numbers, and the chi-square test, ANOSIM analysis, linear discriminant (LEfSe) analysis and the Metastats analysis were used for inter-group comparison. Results:(1) Changes in composition and structure of intestinal microflora. The Shannon index of α diversity of preoperative intestinal microflora in simple obesity patients and obesity combined with diabetes patients was 4.37±0.69 and 4.47±0.85, respectively, showing no significant difference between them ( t=0.28, P>0.05). Results of preoperative LEfSe analysis showed that there were differences in the bacterial abundance of Firmicutes and Bacteroidea between simple obesity patients and obesity combined with diabetes patients. The abundances of Parasutterella in simple obesity patients and obesity combined with diabetes patients was 0.000 113 0(0, 0.004 378 2) and 0.008 464 0(0.001 325 7, 0.034 983 1), respectively, showing a significant difference between them ( Z=2.12, P<0.05). Results of preoperative PCoA analysis showed that the contribution rates of principal component 1, principal component 2 and principal component 3 were 24.98%, 22.24% and 16.33% in simple obesity patients and obesity combined with diabetes patients and results of ANOSIM comparison showed that there was no significant difference in preoperative intestinal microflora between them ( r=?0.11, P>0.05). The Shannon index of α diversity of postoperative intestinal microflora in simple obesity patients and obesity combined with diabetes patients was 4.60±0.65 and 4.66±0.40, respectively, showing no significant difference between them ( t=0.24, P>0.05). Results of postoperative LEfSe analysis showed that there were differences in the bacterial abundance of Bacteroidea, Proteus and Firmicutes between simple obesity patients and obesity combined with diabetes patients. The abundances of Morganella and Coprococcus_2 in simple obesity patients and obesity combined with diabetes patients were 0.000 192 0(0.000 011 9,0.001 569 0), 0(0,0) and 0(0,0), 0.000 054 1(0,0.000 419 0), showing significant differences between them ( Z=2.70, 2.29, P<0.05). (2) Changes of intestinal microflora in simple obesity patients after operation. There were 10 genera of bacteria of intestinal bacteria changing after surgery, including 7 species of bacteria increasing in the Firmicutes and the Proteobacteria as Veillonella, Morganella, Granulicatella, Aeromonas, Streptococcus, Rothia and Megasphaera and the bacteria decreasing in the Firmicutes and the Actinobacteria as Ruminococcus_torques_group, Romboutsia and Erysipelo-trichaceae_UCG-003. Results of LEfSe analysis showed that the bacteria significantly enriched in simple obesity patients before surgery were Ruminococcus_torques_group, Romboutsia and Erysipelotri-chaceae_UCG-003, belonging to Firmicutes, and the bacteria significantly enriched in simple obesity patients after surgery were Rothia, Granulicatella, Enterococcus, Streptococcus, Megasphaera, Veillonella, A eromonas and Morganella, belonging to Actinobacteria, Firmicutes and Proteobacteria. (3) Changes of intestinal microflora in obesity combined with diabetes patients after operation. There were 16 bacteria of intestinal bacteria increasing after surgery, including Streptococcus, Veillonella, Haemophilus, Pluralibacter, Gemella, Lachnospiraceae_NC2004_group, Granulicatella,Aeromonas, uncultured_ bacterium_f_ Saccharimonadaceae, R uminiclostridium_9, Butyricicoccus, Fusobacterium, Anaerotruncus, Fusicateni-bacter, Klebsiella and E ubacterium_eligens_group, which belonged to the Firmicutes and the Proteo-bacteria. Results of LEfSe analysis showed that the bacteria significantly enriched in obesity combined with diabetes patients before surgery were Fusicatenibacter, Tyzzerella_3 and Butyricicoccus, belonging to the Firmicutes, and the bacteria significantly enriched in obesity combined with diabetes patients after surgery were Gemella, Granulicatella, Enterococcus, Streptococcus, Lachnospiraceae_NC2004_group, Eubacterium_eligens_group, Anaerotruncus, Ruminiclostridium_9, Anaeroglobus, Veillonella, Fusobacterium, uncultured_bacterium_f_Saccharimonadaceae, Aeromonas, Klebsiella, Pluralibacter, Proteus and Haemophilus, belonging to the Firmicutes and the Proteobacteria. Conclusions:RYGB can significantly increases the intestinal microflora abundance in simple obesity patients and obesity combined with diabetes patients. The two types of patients have specific changes in intestinal microflora at the genus level.

Objective:To explore the effect of the pharyngeal pressure feedback training on pharyngeal constriction in persons with swallowing disorders caused by brainstem lesions.Methods:Twenty patients with disordered swallowing caused by a brainstem lesion were randomly divided into a control group and an intervention group, each of 10. Both groups received routine swallowing training including oral sensorimotor training, neuromuscular stimulation and balloon catheter dilation, while the intervention group was additionally provided with two weeks of pharyngeal pressure feedback training. Before and after the treatment, both groups were evaluated in terms of the peak pressure of superior and inferior pharyngeal constriction, endoscopically and also using the functional oral intake scale.Results:There was no significant difference between the two groups in any measure before the intervention. Afterward, both groups had improved significantly by all of the measurements, but the average peak upper pharyngeal pressure, the average intake scale score and the endoscopy results of the intervention group were all significantly better than the control group′s averages.Conclusions:Pharyngeal pressure feedback training can significantly relieve swallowing disorders caused by brainstem lesions.

Objective To effectively use the clinical data generated in daily operation and to realize information networking based on the existing resources of radiotherapy department. To improve quality management efficiency in radiotherapy process. Methods The radiotherapy process and required documents were analyzed. The reporting tool Microsoft Report Builder, which is based on SQL database, was applied to design the patient documents by extracting and analyzing a large number of data generated by Aria, the existing network of our radiotherapy department. PDCA Tools was used to analyze the weak links in the process. Reports with quantitative indices have been designed according to corresponding countermeasures, so as to improve quality control level of the process. Results More than one thousand patients were treated in our department since 2020. All patient documents of radiotherapy can be archived and inquired online after registration only once. 13 daily statistical reports, 5 quarters and 3 annual reports were scheduled according to practical demands. The waiting time before radiotherapy was shortened from 16.2 days to 14.8 days after operating the reporting system 3 months later. The staff could master the treatment progress of patients easily and patients who interrupted the treatment were found in time. Conclusion The reporting tools can realize patient information extraction and networked management effectively in radiotherapy process. Staff efficiency of personnel work and communication was improved. The resource allocation was optimized according to the report data in real time, improving the efficiency and quality of radiotherapy. This method is generally applicable and practical to radiotherapy department.

Objective:To determine the viral load of human herpesvirus 6 A (HHV-6A), HHV-6B and chromosomal integrated HHV-6 (ciHHV-6) simultaneously through a triple chip digital PCR (tcdPCR) method for detection of HHV-6A/6B and ribonuclease P-30 (RPP30).Methods:According to optimal reaction conditions of real-time fluorescence quantitative PCR (RT-qPCR) method, the tcdPCR mehod of HHV-6A, HHV-6B and RPP30 was established. The sensitivity of tcdPCR was determined by virus cultures and the specificity of tcdPCR was detected with other herpesviruses. Subsequently, the tcdPCR of HHV-6A, HHV-6B and RPP30 was verified through 127 whole blood samples.Results:The consistency between RT-qPCR and tcdPCR for HHV-6 detection was good (R 2>0.97). And there was no cross-reaction with other herpesviruses. The 14 positive samples could be detected effectively by the tcdPCR of HHV-6A, HHV-6B and RPP30. The lowest detectable viral load of HHV-6A and HHV-6B was 50 copies/ml and 105 copies/ml, respectively. And the ratio of HHV-6/(RPP30/2) in 14 positive samples was less than 1. Conclusions:The tcdPCR has good sensitivity and specificity. And HHV-6 tcdPCR method can quantitatively detect the viral load of HHV-6 infection and the copy number of RPP30, and ciHHV-6 can be judged by ratio of HHV-6/(RPP30/2) in clinical samples.