Objective To analyze the protein expression profile of human glomerular mesangial cells (HMCs) under high glucose and to characterize molecular functions and biological processes. Methods HMCs were divided into high glucose cultured group (30 mmol/L) and normal glucose cultured group (5 mmol/L). The total proteins were extracted after culture for 48 hours. The total proteins of the two groups were separated using two-dimensional fluorescence difference in gel electrophoresis (2-D DIGE) and analyzed using DeCyder 2-D difference analysis software. The differentially expressed proteins were further identified using in-gel digestion with trypsin, of which peptide extracts were prepared for MALDI-TOF-MS analysis. Protein identifications were searched in the NCBI protein database using the Mascot search engine. Results One hundred and forty-seven protein spots whose expression levels were significantly increased or decreased more than 1.5 folds under high glucose were identified. Ninety-six differentially expression protein spots were analyzed by peptide mass fingerprinting and 37 kinds of proteins were identified. The protein spots of phosphatidylethanolamine binding protein 1 (PEBP-1), granulysin,ATP synthase H + transporting mitochondrial FO complex subunit F2 were observed only in high glucose group. The expression of 24 proteins was up-regulated by high glucose, including eosinophil cationic protein, RGS membrane-interacting proteins 16 (MIR16), peptidyl-prolyl cis-trans isomerase, disks large homolog DLG2, breast cancer 2, early onset (BRCA2), Catechol-O-methyltransferase etc. The expression of 5 proteins was down-regulated by high glucose, including O-GlcNAc transferase-interacting protein 106 000 isoform 1, proteasome beta 6 subunit precursor,NEFA-interacting nuclear protein NIP30 etc. Conclusions Expression of 147 proteins in HMCs alters under high glucose. These proteins are involved in the regulation of cytoskeleton, glucose metabolism, cell division, gene transcription, signal transduction, phosphorylation, cell proliferation,apoptosis etc. In-depth analysis of these differentially expressed proteins' function and crosstalk is expected to provide an important experimental basis for clarifying the pathogenesis of diabetic nephropathy.

Objective To investigate The Th1/Th2 and Tc1/Tc2 polarization in the peripheral blood of first degree relatives of pemphigus vulgaris(PV) and healthy control individuals, and to approach the mechanism of the Dsg3-specific autoimmunity in PV. Methods The peripheral blood mononuclear cells (PBMC) from first degree relatives and healthy control was stimulated for72 h with Dsg3 and without Dsg3. Th1/Th2, Tc1/Tc2 was assessed by four-color flow cytometry. Results The mean frequency of Dsg3-spe-cific Th2 cells for PV antibody positive first degree relatives was 10.13%±3.72%, compared with stimula-tion without antigen 7.28%±3.58%, the difference was significant (P<0.05). The percentage Dsg3-spe-cific Th2 was markedly higher in the PV antibody positive first degree relatives group than that in the control group(10.13%±3.72% vs 6.10%±2.82%, P<0.05) , Tc2 was markedly higher also (20.01%± 10.43% v514.91%±8.06%, 20.01%±10.43% vs 9.58%±5.49%, P<0.05). Conclusion When Dsg3 stimulated PBMC were used to stimulate autologous T cells an increased amount of Th2 and Tc2 was observed, it is implied that the imbalance of Th1/Th2, Tc1/Tc2 might play an important role in the initia-tion of PV.

To investigate the neurotoxic effect of human immunodeficiency virus (HIV) gp120 on cultured dorsal root ganglion (DRG)neurons in vitro, dissociated and organotypic mouse embryo's DRG cell culture models were established. Both dissociated and organotypic DRG cultures were treated with HIV gp120 in different concentration (250 pmol/L and 1 nmol/L, respectively, 2 times/7 days). For dissociated DRG cultural cells, microtubule-associated protein 2 (MAP2) immunofluorescent labeling was processed for observing the changes of neuronal cell body and neurites. The change of the ultrastructure in the organotypic cultured DRG was observed by electron microscopy.The difference of the number and length of neurites between the control group and HIV gp120 treated groups were significant (P＜0.001),whereas there was no significant difference in the diameter of neurons between them (P＞0.05). The ultrastructural changes included the decrease or loss of cristae in mitochondria and accumulation of many high densed particles between the microtubules and the neurofilaments by using both the concentrations of HIV gp120 treatment. The present results indicate that HIV gp120 had a directly neurotoxic effect on the cultured DRG neurons, especially more sensitive to mitochondria.

Objective To investigate the effect of berberine on proliferation of neural stem cells(NSCs)induced by hydrogen peroxide(H2O2). Methods NSCs from Sprague-Dawley rats were isolated and purified by suspension culture. Cells were divided into a control group,H2O2group(NSCs exposed to H2O2injury),berberine group(NSCs were incubated with berberine concentrations ranging from 0.5 to 20 μmol/L and exposed to H2O2), and DAPT(a blocker of the Notch signaling pathway)group. Cell viability was evaluated using the Cell Counting Kit-8 assay. Proliferation of NSCs was evaluated by a neurosphere formation assay and Ki67 protein expression. Expression of key proteins in the Notch signaling pathway(including notch1 and hes1)in response to berberine treatment or DAPT(a Notch inhibitor)was determined by Western blotting. Results Cell viability of NSCs was significantly increased by berberine compared with the H2O2group. The neurosphere growth assay showed that 5 or 10 μmol/L berberine increased NSC proliferation. The ratio of Ki67 +/DAPI cells and notch1 and hes1 protein expression increased significantly compared with the H2O2group. Conclusions Berberine treatment upregulates Notch signaling in NSCs,whereas DAPT attenuates these effects. Berberine is a drug that promotes NSC proliferation and exerts a protective effect on NSCs via the Notch signaling pathway.

Objective To investigate the correlation between mild cognitive impairment (PD-MCI) and serum homocysteine (Hcy) in patients with Parkinson's disease.Methods 85 patients with PD (43 with PD-MCI,42 with PDN) as the observation group and 43 healthy elderly patients as the control group.The demographic information,clinical and neuropsychological assessments were conducted and the Hcy of fasting venous blood were detected in all subjects.The I-Icy level and its associated factors were also analyzed in the PD group.Results Serum levels of Hcy in the PD patients((13.53±2.29) μmol/L) were higher than those in the HC group((11.77±1.56) μmol/L),the difference was statistically significant (t=4.57,P ＜0.05).There was significant difference in serum Hcy levels among HC group,PDN group ((12.85 ± 1.88) μmoL/L) and PD-MCI group((14.22±2.47) ±mol/L) (F=16.13,P＜0.01).Hcy level of the PDN group was higher than that of the HC group (P＜0.05),and PD-MCI group was higher than PDN group (P＜0.05).Serum levels of Hcy in patients with PD-MCI were significantly associated with UPDRS-Ⅲ (r=0.305),H-Y (r=0.313),LEDD (r=0.424),MMSE (r =-0.470),MoCA (r=-0.503),duration of connection test B (r =0.617) and semantic fluency tests (r=0.557) (all P＜0.05).Conclusion Elevated serum Hcy levels are expected to be biomarkers for predicting PD-MCI.

Objective:To observe the long-term efficacy and late adverse reactions of simultaneous integrated boost intensity-modulated radiotherapy (SIB-IMRT) for mediastinal lymph node recurrence after radical surgery for esophageal squamous cell carcinoma (ESCC).Methods:A total of 20 ESCC patients with mediastinal lymph node recurrence (≤5) after radical surgery admitted to Department of Radiotherapy, Shanghai Ruijin Hospital between June 2019 and December 2021 were enrolled in this prospective study. Among them, 10 patients were enrolled in phase I study and 10 patients in phase II study. Four, 3 and 13 patients received three different doses of SIB-IMRT at 58.8 Gy/28 fractions, 64.4 Gy/28 fractions and 70.0 Gy/28 fractions for recurrent lesions, respectively. The overall survival (OS) rate, local control rate (LCR) and progression-free survival (PFS) were calculated by Kaplan-Meier analysis. Adverse reactions were also analyzed.Results:The most common sites of recurrence were 2R and 4 L, accounting for 35% and 25%, respectively. The median follow-up time was 32 months. For patients who received salvage chemoradiation after relapse, the 1-, 2- and 3-year OS rates were 100%, 88% and 78%, the 1-, 2- and 3-year PFS rates were 85%, 78% and 78%, respectively. The most common hematological toxicities were leukocytopenia and anemia. The most common nonhematological toxicity was esophagitis. However, no grade 3 or above esophagitis, pneumonia and cardiotoxicity were found. Three patients who received SIB-IMRT at 58.8 Gy/28 fractions died of distant metastases at 2 years after treatment, and 1 patient who received SIB-IMRT at 70.0 Gy/28 fractions died of distant metastases at 16 months after treatment.Conclusion:Salvage chemoradiotherapy using SIB-IMRT is efficacious and safe for mediastinal lymph node recurrence in ESCC patients after radical resection.

BACKGROUND:Currently, hematopoietic stem celltransplantation mainly depends on unrelated donors. Mental state of the unrelated donors is very important to ensure the successful celltransplantation. OBJECTIVE:To compare mental and physical health status of relative and unrelated donors during the hematopoietic stem cellcol ection. METHODS:We compared the mental (Symptom Checklist-90) and physical (temperature, breath, pulse, and blood pressure) health status of relative and unrelated donors at admission, 1 day before cellcol ection, and 1-2 days after cellcol ection.RESULTS AND CONCLUSION:At admission, there was no difference in the mental health status of relative and unrelated donors (P>0.05), while the scores on Symptom Checklist-90 were significantly higher in the unrelated donors than the relative donors, including total score, forced, depression, anxiety, hostility, and fear (P<0.05). The physical signs were steady in the unrelated and relative donors, but the difference in breath and systolic blood pressure was of great significance before and after cellcol ection in the two groups. These findings indicate that during cellcol ection, the unrelated donors exhibit heavier mental load than the relative donors, and psychological counseling and health guidance are necessary.

ObjectiveTo investigate the role and prognostic value of matrix metalloproteinase-7 （MMP7） in the migration and immune cell infiltration of hepatocellular carcinoma. MethodsMMP7_siRNA for downregulating the target gene MMP7 and pMMP7 for upregulating MMP7 were constructed and were used to transfect hepatocellular carcinoma cell line （MHCC97H）. RT-qPCR and Western Blot were used to measure the mRNA and protein expression levels of the target gene in cells. Scanning electron microscopy and Transwell assay were used to observe the changes in cell pseudopodia and migration ability， and bioinformatics methods were used to investigate the correlation of MMP7 with immune cells and immune infiltration score in TCGA and TIMER databases in patients with hepatocellular carcinoma， as well as the association between MMP7 and the prognosis of patients with hepatocellular carcinoma. The Spearman method was used for correlation analysis. Sanger Box online tool was used to assess the value of MMP7 in the overall survival curve and disease-specific survival of hepatocellular carcinoma. The Kaplan-Meier method was used to plot survival curves， and the Log-rank test was used for comparison of prognosis between different samples. ResultsAfter MHCC97H cells were transfected with MMP7_siRNA or pMMP7， there was a significant reduction or increase in the expression of the target gene MMP7； after downregulation of MMP7， there were significant reductions in the number and length of the pseudopodia， while after MMP7 overexpression， there were significant increases in the number and length of filopodia with radial arrangement. The Transwell chamber assay showed that MMP7_siRNA2 significantly reduced the migration ability of cells （P<0.05）， and there was a significant increase in migration ability after pMMP7 transfection. The expression of MMP7 was significantly correlated with B lymphocytes （r=0.37， P<0.05）， CD4⁺ T lymphocytes （r=0.40， P<0.05）， neutrophils （r=0.49， P<0.05）， macrophages （r=0.49， P<0.05）， and dendritic cells （r=0.47， P<0.05）. In the TCGA database， the patients with hepatocellular carcinoma were divided into MMP7 high expression group with 267 patients and MMP7 low expression group with 146 patients based on overall survival， and the results showed that the MMP7 high expression group had a significantly shorter overall survival time than the MMP7 low expression group （P<0.05）； based on the disease-specific survival time， the patients were divided into MMP7 high expression group with 257 patients and MMP7 low expression group with 145 patients， and the analysis showed that the MMP7 high expression group also had a significantly shorter disease-specific survival time than the MMP7 low expression group （P<0.05）. ConclusionMMP7 promotes the migration of hepatocellular carcinoma cells and plays a major role in immune cell infiltration， and the expression of MMP7 is also significantly associated with the prognosis of hepatocellular carcinoma.