Objective:To investigate the effect of Xuebijing injection against blood-brain barrier(BBB)damage in the mice with anti-N-methyl-D-aspartate receptor(NMDAR)encephalitis,and to elucidate its regulatory effect on the imbalance of helper T cells 17(Th17)/regulatory T cells(Treg).Methods:The active immunization models of anti-NMDAR encephalitis in the mice were established using glutamate receptor N1 subunit(GluN1)356-385 antigen peptide,and the serum anti-NMDAR immunoglobulin G(IgG)antibody levels were detected by enzyme-linked immunosorbent assay(ELISA).The healthy mice without modeling were served as control group,and the mice with successful modeling were randomly divided into model group,low dose of Xuebijing injection(XBJ-L)group,and high dose of Xuebijing injection(XBJ-H)group,with 10 mice in each group.After modeling,the mice in XBJ-L and XBJ-H groups were intraperitoneally injected with 5 and 10 mL·kg-1 Xuebijing injection,respectively.The Longa score was used to assess the neurological impairment of the mice in various groups;evans blue(EB)staining was used to determine the BBB permeability;immunofluorescence staining was used to detect the expressions of zonula occludens 1(ZO-1)and Occludin in cerebral cortex of the mice in various groups;Western blotting method was used to determine the expression levels of ZO-1,Occludin,Claudin-5,and neuron-specific nuclear protein(NeuN)in cerebral cortex of the mice in various groups;ELISA method was used to determine the levels of Th17-and Treg-related cytokines including interleukin(IL)-17,IL-22,and IL-10 in serum of the mice;flow cytometry was used to determine the percentages of Th17 and Treg cells in peripheral blood of the mice in various groups,and the Th17/Treg ratio was calculated.Results:The serum of the mice induced with the GluN1 356-385 antigen peptide was positive for NMDAR IgG antibodies,indicating that the models were successfully established.Compared with control group,the neurological impairment score of the mice in model group was significantly increased(P＜0.05),and the EB level in brain tissue was significantly increased(P＜0.05);the fluorescence staining intensities of ZO-1 and Occludin in the cerebral cortex were decreased,and the expression levels of ZO-1,Occludin,Claudin-5,and NeuN proteins in the cerebral cortex were significantly decreased(P＜0.05);the serum levels of IL-17 and IL-22 were significantly increased(P＜0.05),while the IL-10 level was significantly decreased(P＜0.05);the percentage of Th17 cells in peripheral blood was significantly increased(P＜0.05),while the percentage of Treg cells was significantly decreased(P＜0.05),and the Th17/Treg ratio was significantly increased(P＜0.05).Compared with model group,the neurological impairment scores of the mice in XBJ-L and XBJ-H groups were significantly decreased(P＜0.05),the EB levels in brain tissue were significantly decreased(P＜0.05),the fluorescence staining intensities of ZO-1 and Occludin in cerebral cortex were increased,and the expression levels of ZO-1,Occludin,Claudin-5,and NeuN proteins were significantly increased(P＜0.05);the levels of IL-17 and IL-22 in serum were significantly decreased(P＜0.05),and the level of IL-10 was significantly increased(P＜0.05);the percentages of Th17 cells in peripheral blood were significantly decreased(P＜0.05),the percentages of Treg cells were significantly increased(P＜0.05),and the Th17/Treg ratios were significantly decreased(P＜0.05).Compared with XBJ-L group,the neurological function injury score of the mice in XBJ-H group was significantly decreased(P＜0.05),the EB level in brain tissue was significantly decreased(P＜0.05);the fluorescence staining intensities of ZO-1 and Occludin in the cerebral cortex were increased,and the expression levels of ZO-1,Occludin,Claudin-5,and NeuN proteins were significantly increased(P＜0.05);the serum levels of IL-17 and IL-22 were significantly decreased(P＜0.05),and the level of IL-10 was significantly increased(P＜0.05);the percentage of Th17 cells in peripheral blood was significantly decreased(P＜0.05),the percentage of Treg cells was significantly increased(P＜0.05),and the Th17/Treg ratio was significantly decreased(P＜0.05).Conclusion:Xuebijing injection can improve BBB injury,regulate Th17/Treg balance,and thereby alleviate the neurological functional damage in anti-NMDAR encephalitis.

Objective:To investigate the differences in the expression profiles of cyclic RNA (circRNA) in peripheral blood mononuclear cells (PBMCs) of rheumatoid arthritis (RA) and its clinical significance.Methods:Venous blood were collected from 4 patients with RA (group T) and 4 healthy subjects (group C). The expression profiles of circRNA in PBMCs of the two groups were detected by Arraystar circRNA microarray, and the differentially expressed circRNA was analyzed by clustering analysis. The binding sites for interaction between differentially expressed circRNA and miRNA were predicted, and functional analysis such as geneontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis was performed. quantitative real-time polymerase chain reaction (RT-qPCR) was used to verify the expression of partially differentially expressed circRNA in the two groups of PBMCs, and a circRNA-miRNA-mRNA regulatory network (ceRNA network) was constructed for the target circRNA with significantly differential expression. A receiver operating characteristic curve [receiver operating characteristic curve (ROC)] was established to analyze the potential diagnostic value of target circRNA. SPSS Statistics 23.0 and Graphpad Prism 8.0 were used to analyze the data, and the independent t test was used to analyze the difference between the two groups. Results:① Microarray results showed that, compared with group C, a total of 399 [fold of difference (FC)>1.5, and P<0.05] circRNA were abnormally expressed in PBMCs of group T; including 149 up-regulated and 250 down-regulated. ② Bioinformatics analysis: The prediction of the binding site of circRNA and miRNA suggested that the differentially expressed circRNA in RA might affect the inflammatory response by targeting miR-140-5p, miR-338-5p, and miR-9-5p. GO analysis showed that the differentially expressed circRNA was mainly involved in the intimal-binding organelles, protein metabolism and binding, etc. KEGG pathway analysis showed that most of the involved pathways were related to infection and human immune dysregulation. ③ The results of multi-sample RT-qPCR validation showed that the expression level of hsa_circRNA_009012 in group T was significantly higher than that in group C ( t=-4.417, P<0.01), the expression level of hsa_circRNA_101328 was significantly lower than that in group C ( t=-1.042, P<0.01), and the expression of hsa_circRNA_058230 had no significant change ( t=4.691, P>0.05). ④ ROC curve analysis indicated that hsa_circRNA_009012 had potential value in the diagnosis of RA [area under curve=0.96]. Conclusion:The expression of circRNA in PBMCs of patients with RA is imbalanced, and it may participate in the regulation of the development of RA. Among them, hsa_circRNA_009012 is expected to become a new biological marker for the diagnosis and treatment of RA.

Objective:To clarify the correlation of the expression of glia maturation factor-β (GMF-β) with Ki-67 in astrocytoma, and to investigate the prognostic implications of combined detection of GMF-β and Ki-67.Methods:One hundred and forty human astrocytoma samples (WHO Ⅱ-Ⅳ grade) were collected at Southwest Hospital, Army Medical University (the Third Military Medical University), China from 2006 to 2009. Clinicopathological information and 3-year follow-up data were collected. Expression of GMF-β and Ki-67 was detected by single and double immunohistochemical staining, then the association of GMF-β expression with Ki-67 and its significance in prognostic evaluation of astrocytoma were statistically analyzed.Results:GMF-β expression in astrocytoma cells was correlated to both tumor grade and Ki-67 (both P<0.05); Kaplan-Meier survival analysis showed that GMF-β and Ki-67 expression were negatively correlated to the 3 year-survival rates, respectively (both P<0.01). Further analysis demonstrated that the two factors were co-influenced on survival, showing a trend of "GMF-β low Ki-67 low>GMF-β high Ki-67 low>GMF-β low Ki-67 high>GMF-β high Ki-67 high" in 3-year survival rate with significant intergroup differences ( P<0.05, P<0.01). Conclusions:GMF-β expression is positively associated with Ki-67 in astrocytoma. Combined detection of GMF-β and Ki-67 can predict prognosis of patients with glioma.

Animals ; Cdc20 Proteins/metabolism* ; Cell Line ; Embryo, Mammalian/embryology* ; Embryonic Development ; Female ; Infertility, Female/metabolism* ; Mice ; Mutation ; Oocytes/metabolism*

OBJECTIVE: To investigate high-salt exposure-induced polarization of mononuclear macrophages and the changes in proliferation and phenotypic transformation of renal fibroblasts in a co-culture system. METHODS: Cultured mononuclear macrophages were exposed to high salt (161 mmol/L Na +) for 2 h and the surface markers of M0, M1 and M2-type macrophages were detected with RT-qPCR. The culture medium of the macrophages in normal and high-salt groups was collected for detection of the mRNA and protein levels of IL-6 and TGF-β1 using RT-qPCR and ELISA. A co-culture system of high salt-exposed macrophages and renal fibroblasts (NRK-49F) was established using a Transwell chamber, and the changes in proliferation and migration of NRK-49F cells were examined using EdU assay and Transwell assay, respectively. Western blotting was performed to detect the expressions of collagen I, collagen III and collagen α-SMA in NRK-49F cells. RESULTS: The high salt-exposed macrophages showed significantly increased mRNA levels of M2-type macrophage surface markers mannose receptor and arginase ( CONCLUSIONS High-salt exposure induces polarization of mononuclear macrophages into M2-type macrophages and promotes secretion of IL-6 and TGF-β1 by the macrophages to induce the proliferation and phenotypic transformation of NRK-49F cells.
Cell Proliferation ; Coculture Techniques ; Fibroblasts ; Kidney ; Macrophages ; Transforming Growth Factor beta1/genetics*

Objective: To investigate the safety and efficacy of surgical treatment for chronic radiation intestinal injury. Methods: A descriptive cohort study was performed. Clinical data of 73 patients with definite radiation history and diagnosed clinically as chronic radiation intestinal injury, undergoing operation at Department of Colorectal Surgery, the Sixth Affiliated Hospital of Sun Yat-sen University from January 1, 2012 to February 28, 2019, were reviewed and analyzed retrospectively. Patients did not undergo operation or only received adhesiolysis were excluded. All the patients had preoperative examination and overall evaluation of the disease. According to severity of intestinal obstruction and patients′ diet, corresponding nutritional support and conservative treatment were given. Surgical methods: The one-stage bowel resection and anastomosis was the first choice for surgical treatment of chronic radiation intestinal injury. Patients with poor nutritional condition were given enterostomy and postoperative enteral nutrition and second-stage stoma closure and intestinal anastomosis if nutritional condition improved. For those who were unable to perform stoma closure, a permanent stoma should be performed. Patients with severe abdominal adhesion which was difficult to separate, enterostomy or bypass surgery after adhesiolysis would be the surgical choice. For patients with tumor metastasis or recurrence, enterostomy or bypass surgery should be selected. Observation parameters: the overall and major (Clavien-Dindo grades III to V) postoperative complication within 30 days after surgery or during hospitalization; mortality within postoperative 30 days; postoperative hospital stay; time to postoperative recovery of enteral nutrition; time to removal of drainage tube. Results: Of the 73 patients who had been enrolled in this study, 10 were male and 63 were female with median age of 54 (range, 34-80) years. Preoperative evaluation showed that 61 patients had intestinal stenosis, 63 had intestinal obstruction, 11 had intestinal perforation, 20 had intestinal fistula, 3 had intestinal bleeding, and 6 had abdominal abscess, of whom 64(87.7%) patients had multiple complications. Tumor recurrence or metastasis was found in 15 patients. A total of 65(89.0%) patients received preoperative nutritional support, of whom 35 received total parenteral nutrition and 30 received partial parenteral nutrition. The median preoperative nutritional support duration was 8.5 (range, 6.0-16.2) days. The rate of one-stage intestine resection was 69.9% (51/73), and one-stage enterostomy was 23.3% (17/73). In the 51 patients undergoing bowel resection, the average length of resected bowel was (50.3±49.1) cm. Among the 45 patients with intestinal anastomosis, 4 underwent manual anastomosis and 41 underwent stapled anastomosis; 36 underwent side-to-side anastomosis, 5 underwent end-to-side anastomosis, and 4 underwent end-to-end anastomosis. Eighty postoperative complications occurred in 39 patients and the overall postoperative complication rate was 53.4% (39/73), including 39 moderate to severe complications (Clavien-Dindo grade III-V) in 20 patients (27.4%, 20/73) and postoperative anastomotic leakage in 2 patients (2.7%, 2/73). The mortality within postoperative 30 days was 2.7% (2/73); both patients died of abdominal infection, septic shock, and multiple organ failure caused by anastomotic leakage. The median postoperative hospital stay was 13 (11, 23) days, the postoperative enteral nutrition time was (7.2±6.9) days and the postoperative drainage tube removal time was (6.3±4.2) days. Conclusions Surgical treatment, especially one-stage anastomosis, is safe and feasible for chronic radiation intestine injury. Defining the extent of bowel resection, rational selection of the anatomic position of the anastomosis and perioperative nutritional support treatment are the key to reduce postoperative complications.

Objective To study the feasibility,efficacy and safety of ultrasound-guided percutaneous radiofrequency ablation (RFA) in the treatment of hepatocellular carcinoma (HCC) in the caudate lobe.Methods From November 2006 to June 2017,31 patients with 31 HCCs located in the caudate lobe were treated with percutaneous RFA at the First Affiliated Hospital of Sun Yat-sen University.The treatment efficacy,complications,and the local tumor progression (LTP),disease-free survival (DFS) and overall survival (OS) rates were analyzed.Results Residual tumors were detected in 5 patients after the first treatment.Complete necrosis was achieved in all the patients after the second treatment.The mean number of ablation sessions was 1.16±0.37.At a follow-up period which ranged from 3 to 65 months,19 patients had died,10 patients were still alive,and 2 patients were lost to follow-up.The 1-,2-,3-,and 5-years OS rates were 78.4％,48.5％,12.1％ and 12.1％,respectively.On follow-up,9 caudate lobe HCC lesions were detected to have LTP.The 1-,2-,and 3-years LTP rates were 21.5％,41.6％ and 41.6％,respectively;while the 1-,2-,and 3-years DFS rates were 22.3％,11.2％ and 11.2％,respectively.Ablationrelated complications were detected in 4 patients.Conclusions Ultrasound-guided percutaneous RFA was safe and effective for patients with HCC in the caudate lobe.These patients should be followed-up closely to detect LTP.

Objective To retrospectively compare the safety and local efficacy of multiple-electrode switching based radiofrequency ablation (RFA) and the conventional RFA in treating single early-stage hepatocellular carcinoma (HCC).Methods A total of 82 patients with single early-stage HCC received either RFA with a multiple-electrode switching system (n =43) or conventional RFA (n =39) as the first-line treatment.The rate of initial local complete response,major complications and local tumor progression (LTP) were compared between two groups.Results The total ablation time was significantly shorter in the switching-RFA group [(16.7 ± 3.4) mins] than in the conventional RFA group [(29.8 ± 10.4) mins] (P ＜ 0.05).The rate of initial local complete response was 100％ (43/43) in the switching-RFA group and 94.9％ (37/39) in the conventional RFA group (P ＞0.05).After a mean follow-up period of (26.4 ± 21.8)months (ranging 3.0-91.6 months),the rates of LTP in the switching-RFA group and conventional RFA group were 16.3％ (7/43) and 17.9％ (7/39),respectively.The LTP rates in two groups were 16.1％ versus 11.2％ atyear1,and20.5％ versus 20.6％ at year2 (P=0.666).Conclusions The multiple-electrode switching based RFA is safe and effective with shorter ablation time in treating single early-stage hepatocellular carcinoma.

AIM: To investigate the primary culture method for coronary artery smooth muscle cells (CASMCs), and to establish the endoplasmic reticulum stress ( ERS) model in CASMCs of SD rats.METHODS:CASMCs were cultured by tissue explant method .The morphological characteristics were observed under optical micro-scope.The marker proteins of CASMCs , including α-SMA and SM-MHC, were identified by immunofluorescence tech-nique.The protein expression levels of BiP and CHOP , the marker molecules of ERS, were determined by Western blot . RESULTS:The spindle-shaped CASMCs climbed out from the edge of coronary artery tissues after 6 d, and formed the typical hill and valleygrowth pattern of CASMCs at 9~10 d.The result of immunofluorescence technique showed that α-SMA and SM-MHC were positively expressed .The results of Western blot showed that the protein expression of BiP and CHOP in TG ( 1 and 2 μmol/L ) treatment groups was increased compared with control group .Compared with control group, the protein expression of BiP and CHOP was significantly increased after 1 μmol/L TG treatment for 24 and 48 h. CONCLUSION:CASMCs can be successfully cultured by tissue explant method .ERS model of CASMCs was established by 1 μmol/L TG treatment for 24 h.

Objective To investigate the role of plasma matrix metalloproteinase-9 (MMP-9) and nuclear factor kappa B (NF-κB) in the pathogenesis of obstructive sleep apnea hypopnea syndrome (OSAHS) complicated with hypertension.Methods The patients were selected in the respiratory medicine and neurology of our hospital from January 2014 to June 2014.Polysomnography(PSG) monitoring was employed to assess the patient.All patients were divided into 3 groups,including snoring group(23 cases),OSAHS group(32 cases),OSAHS with hypertension group(31 cases).And then according to the hypertension stage,OSAHS with hypertension group was divided into 3 groups,including OSAHS with stage 1 hypertension group(9 cases),OSAHS with stage 2 hypertension group(9 cases),OSAHS with stage 3 hypertension group(13 cases).The plasma MMP-9 was measured by ELISA.The mRNA expression levels of MMP-9 and NF-κB were detected by real-time quantitative PCR.Results The levels of MMP-9 and MMP-9 mRNA and NF-κB mRNA in OSAHS with hypertension group were significantly higher than those of snoring group and OSAHS group and the differences were statistically significant(P<0.05).The expression of MMP-9,the MMP-9 and NF-κB mRNA expression of monocytes in OSAHS with stage 3 hypertension group was significantly higher than those of OSAHS with stage 1 hypertension group and OSAHS with stage 2 hypertension group and the differences were statistically significant(P<0.05).The plasma MMP-9 was positively correlated with AHI and oxygen desaturation index and negatively correlated with LSaO2(P<0.05).The plasma MMP-9 was positively correlated with the MMP-9 mRNA expression(P<0.05).The MMP-9 mRNA expression was positively correlated with the NF-κB mRNA expression(P<0.05).Conclusion The plasma concentrations of MMP-9,the MMP-9 and NF-κB mRNA expression of monocytes in OSAHS with hypertension patients was significantly high,which is associated with disease severity and degree of hypoxia.The MMP-9 might play an important role in the pathogenesis of OSAHS with hypertension and be regulated by NF-κB pathway.