OBJECTIVE:To study the analgesic effect and mechanism of Gutongtie paste on model rats with formaldehyde-in-duced pain. METHODS:60 SD rats were randomly divided into blank group,model group,Gutongtie paste low-dose,medi-um-dose,high-dose groups(0.594,1.188. 2.376 g/paste,containing crude drug 0.48,0.96,1.92 g)and prednisone acetate group (ig,0.0054 g/kg,external bonding matrix). Model rats with pain was induced by formaldehyde method and immediately adminis-trated after modeling. Electronic tenderness instrument was adopted to determine the pain threshold of rats'ankle joint after adminis-tration of 1,2,3,4,6 h. After 6 h,blood sample 0.3 mL was taken from abdominal aorta then rats were sacrificed. Enzyme linked immunosorbent assay (ELISA) was conducted to determine the β-endorphin (β-EP),prostaglandin E2 (PGE2) contents;spectrophotometry was used to determine nitric oxide(NO)content in rats'serum and inflammatory tissue;and radioimmunoassay was adopted to detect the substance P content in rats'serum,inflammatory tissue and brain tissue. RESULTS:Compared with be-fore modeling,pain thresholds in model group at each period were significantly decreased (P<0.05 or P<0.01). Compared with blank group,PGE2,NO of rats,substance P content in inflammatory tissue and brain tissue in model group were significantly in-creased (P<0.05 or P<0.01). Compared with model group,pain thresholds in Gutongtie paste groups at corresponding time points were increased,PGE2 and substance P contents in inflammatory tissue and brain tissue were decreased (P<0.05 or P<0.01);β-EP and NO contents in serum in Gutongtie paste medium-dose,high-dose groups(P<0.05 or P<0.01),NO contents in serum in Gutongtie paste high-dose group were decreased(P<0.05). CONCLUSIONS:Gutongtie paste has a certain analgesic and anti-inflammatory effect,and the mechanism may be related to reducing PGE2, NO, substance P contents, increasing β-EP content.

The aim of this study was to investigate the effects of CD4(+)CD25(+) regulatory T cells (Treg) on allogeneic hematopoietic stem cell transplantation (HSCT) in sensitized mice so as to provide experimental evidence for clinical treatment of allogeneic HSCT rejection in sensitized recipients. The BALB/c mice were divided into 5 groups: group A - mice were sensitized with injection of splenocytes; group B - mice were sensitized with splenocytes and treated with >5×10(5) Treg on day 7 before transplantation; group C - mice were sensitized with splenocytes and treated with 5×10(5) Treg on day 13 and 7 before transplantation; group D - mice were not sensitized, but treated with equal volume of PBS as control; group E - blank control. Each group had 15 mice. On day 0 of transplantation, mice in each group were irradiated lethally with 8 Gy by linear accelerator, and the bone marrow cells of C57BL/6 labeled by fluorescence staining were intravenously injected via the tail vein. The fluorescent cells in peripheral blood and organ tissue were detected by flow cytometry on different time points for homing assessment. Survival rates and hematopoietic reconstitution were also recorded and monitored. The results showed that on 12 and 24 hours after transplantation, as compared with the sensitized group, the number of fluorescence homing cells in different tissue of the applied Treg groups increased significantly and the differences were statistically significant (P < 0.05). The mice in sensitized group and blank control group all died on the 6-13 day, whereas the median survival time of mice in applied Treg once and twice were 15 days and 16 days respectively. Comparing with sensitized group, the difference was statistically significant (P < 0.001), but there was no significant difference between these two groups applied regulatory T cell (P > 0.05). It is concluded that applying Treg can induce immune tolerance of sensitized recipient to allogeneic HSCT and inhibit immune destruction and prolong the survival time, but can not induce full immune tolerance and at last sensitized mice died of rejection of hematopoietic stem cells.
Animals ; Graft vs Host Disease ; etiology ; Hematopoietic Stem Cell Transplantation ; methods ; Immune Tolerance ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; T-Lymphocytes, Regulatory ; immunology ; Transplantation, Homologous

This study was aimed to explore the effects of blocking B7/CD28 and CD40/CD154 co-stimulatory signals on immune function of sensitized mice', and provide the evidences of acquired immune tolerance for allogeneic bone marrow transplantation. The mice sensitized on 7 day before transplant were divided into 4 groups: (1)CTLA4Ig+ anti-CD154 isotype control IgG; (2)anti-CD154 +CTLA4Ig isotype control IgG; (3)CTLA4Ig and anti-CD154; (4)isotype control IgG of CTLA4Ig and anti-CD154. CTLA4Ig and anti-CD154 used in normal BALB/c mice as isotype control IgG. Each mouse in all groups received CTLA4Ig and anti-CD154 (or corresponding isotype control IgG) 500 µg respectively, and was injected via tail vein on 7 day before transplant. There were 5 mice in each group. The mice were sacrificed on day 0, then the number of CD19(+)CD69(+)B cells, CD44(high)/CD62L(high) and CD44(high)/CD62L(low)/- T cells were measured by flow cytometry. Changes of cytokines and sensitized antibody were tested by ELISA or flow cytometry. The results showed that the numbers of CD19(+)CD69(+)B cells were significantly increased in comparison with the normal group (P < 0.01) , whereas the numbers of cells were significantly decreased when blocking B7/CD28 or /and CD40/CD154 co-stimulatory signals (P < 0.01) . Blocking these 2 signals together displayed a synergistic effect (P < 0.01) . The central memory and effector T cells were defined as CD44(high)/CD62L(high) and CD44(high)/CD62L(low)/- respectively, those increased significantly after sensitized in comparison with those in normal group, whereas their numbers decreased when blocking B7/CD28 or/and CD40/CD154 co-stimulatory signals. Blocking these two signals together, displayed a synergistic effect (P < 0.01). Cytokines, IgG and IgM in all groups were not significantly different. Sensitizing antibody test showed that the fluorescence intensity of sensitized group significantly increased as compared with normal group, whereas fluorescence intensity of CTLA4Ig or/and anti-CD154 treated groups significantly decreased as compared with sensitized group (P < 0.01) . It is concluded that blocking the B7/CD28 or/and CD40/CD154 co-stimulatory signal can inhibit the cellular and humoral immune function, whereas blocking these two signals together displays a synergistic effect.
Animals ; B7-1 Antigen ; metabolism ; Bone Marrow Transplantation ; CD28 Antigens ; metabolism ; CD40 Antigens ; metabolism ; CD40 Ligand ; metabolism ; Immune Tolerance ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Signal Transduction ; Transplantation, Homologous

OBJECTIVE: To examine the correlation of CCBE1 expression in adjacent tissues of tongue squamous cell carcinoma (TSCC) with pericancerous lymphatic vessel proliferation, cervical lymph node metastasis and survival outcomes of the patients. METHODS: Lymphatic vessel density was quantified in pericancerous tissue sections of 44 cases of cT_1-2N₀ TSCC using D2-40 as the lymphatic vessel endothelial marker for calibration and counting of the lymphatic vessels. Of these 44 cases, 22 showed a relatively low lymphatic vessel density (group A) and the other 22 had a high lymphatic vessel density (group B), and the expression levels of CCBE1 in the adjacent tissues determined using immunohistochemistry, immunofluorescence assay and Western blotting were compared between the two groups. The expression level of CCBE1 was also measured in another 90 patients with TSCC using immunohistochemistry, and all the patients were followed up for their survival outcomes. RESULTS: Immunohistochemistry and Western blotting showed a significantly lower rate of high CCBE1 expression in group A than in group B (P < 0.05). Immunofluorescence assay showed co-localization of CCBE1 and D2-40 in the adjacent tissues of TSCC. In the 90 TSCC patients with complete follow-up data, a high expression of CCBE1 was found to correlate with lymph node metastasis and a poor 5-year survival outcomes of the patients (P < 0.05). CONCLUSION A high expression of CCBE1 in the adjacent tissues of TSCC is closely related with pericancerous lymphatic vessel proliferation, cervical lymph node metastasis and a poor 5-year survival of the patients, suggesting the value of CCBE1 as a potential prognostic predictor for TSCC.
Humans ; Tongue Neoplasms/pathology* ; Carcinoma, Squamous Cell/metabolism* ; Lymphatic Metastasis ; Prognosis ; Lymphatic Vessels/pathology* ; Cell Proliferation ; Tongue/pathology* ; Calcium-Binding Proteins/metabolism* ; Tumor Suppressor Proteins/metabolism*

Recent clinical trials have shown that electrical stimulation has beneficial effects in obstructive sleep apnea syndrome (OSAS). The purpose of this study was to evaluate the efficacy of electrical stimulation therapy for OSAS with a meta-analysis. The meta-analysis of all relative studies was performed through searching international literature, including PUBMED, CNKI, and EMBASE databases. This literature analysis compared all patients undergoing electrical stimulation therapy with respect to the respiratory disturbance index (RDI) and changes in sleep structure. Six studies were selected involving a total of 91 patients. The meta-analysis indicated that electrical stimulation therapy reduced RDI, longest apnea time, and improved the minimum SaO2. Based on the evidence found, electrical stimulation may be a potential therapy for OSAS, warranting further clinical trials.
Electric Stimulation Therapy ; methods ; Humans ; Respiratory Mechanics ; physiology ; Sleep ; physiology ; Sleep Apnea, Obstructive ; physiopathology ; therapy

OBJECTIVETo study the effect of hepatocyte growth factor on the proliferation of dental pulp cell.

METHODSThe 4th generation dental pulp cell cultured in vitro was used as target cell. 1- 200 microg/L hepatocyte growth factor was added in the test group while the pure cell culture DMEM as control. The MTT method and flowcytometry were applied to assay the proliferation and cell cycle of dental pulp cell of different groups.

RESULTS1-200 microg/L hepatocyte growth factor showed promoting effect to the proliferation of pulp cell since the 5th day (P < 0.05). 100 microg/L was found to be the optimal concentration. Also on the 5th day, 100 microg/L hepatocyte growth factor decreased the G1 subcycle and increased the S subcycle of dental pulp cell ( P < 0.05). While on the 3rd day, it had no effect on the cell cycle.

CONCLUSIONHepatocyte growth factor had positive effect on the proliferation of dental pulp cell, with 100 microg/L as the optimal concentration.

Objective： The circadian rhythm of DNA synthesis in nasopharyngeal carcinoma (NPC) cells was investigated so that to provide necessary data for making clinical chrono-chemotherapy schedule of NPC. Methods： Fifteen BALB/c mice were synchronized with an alternative lighting regimen with 12 hours for light and 12 hours for dark (12∶ 12 LD) at least 3 weeks. Human nasopharyngeal poorly differentiated squamous cell carcinoma(CNE-2) cells were implanted into both flanks of each mouse. Ten days after transplantation, sampling from tumor was conducted in the 3, 9, 15, 21 hours after light onset (HALO). Single cell suspension was obtained and stained with propidium iodide. The cellular DNA content was measured by flow cytometry. Data were documented by ANOVA and Cosinor analysis. Results： The proportion of tumor cells in Phage G1,S,G2-M varied according to circadian time with statistical significance. The distribution curves of Phage G1 and Phage G2-M fit for Cosinor changes. Conclusion： DNA synthesis of human nasopharyngeal poorly differentiated squamous cell carcinoma(CNE-2) cells trausplanted in nude mice varies according to circadian rhythm.

In order to investigate the relationship between gut microbiota and type 2 diabetic erectile dysfunction (T2DED),we analyzed the characteristics of gut microbiota in the Sprague-Dawley (SD) rats with T2DED.Thirty-five SD rats were randomly divided into two groups:control group (n=15)with normal diet,and experimental group (n=20) with construction of T2D model.Faecal and serum samples were collected at 2nd and 8th week after establishment of T2D model,respectively.Faecal samples were used for analysis of gut microbiota,and serum samples for detection of trimethylamine N-oxide (TMAO),lipopolysaccharide (LPS),and inflammatory factors like interleukin-1 (IL-1),IL-2,IL-10,and monocyte chemoattractantprotein-1 (MCP-1).The main compositions of gut microbiota were Bacteroidetes,Proteobacteria and Firmicutes at the phylum level,and Oscillospira,Allobaculum,Bacteroides,Ruminococcus,SMB53,Prevotella,Coprococcus,Sutterella and Blautia at the genus level with relatively higher abundance in all SD rats.The relative abundance of Enterococcus,Corynebacterium,Aerococcus,Facklamia (opportunistic pathogens in most case) increased,and that ofAllobaculum,Bifidobacterium,Eubacterium,Anaerotruncus (beneficial bacteria) decreased in T2DED group as compared with that at 2nd week after establishment of T2D model (T2D2 group).The serum contents of TMAO,LPS,IL-1,IL-2,IL-10 and MCP-1 in T2DED group were significantly higher than those in control group.The gut microbiota of T2DED rats was inhibited.The gut microbiota of T2DED rats had changed,as the relative abundance of beneficial bacterium was decreased while that of opportunistic pathogens was increased.The variations of gut microbiota might lead to inflammation and prompt the emergence of erectile dysfunction in the rats with T2D.TMAO might play an important role in the formation of T2DED.

This research was aimed to explore the effects of blocking B7/CD28 and CD40/CD154 co-stimulatory signals on engraftment of hematopoietic stem cell in the sensitized recipient so as to provide the experimental evidence for the treatment of sensitized recipient's immune rejection after clinical allogeneic hematopoietic stem cell transplantation (HSCT). The BALB/c mice were divided into 4 groups: (1)mice sensitized on 7 day before transplant; (2)mice were sensitized on 7 day before transplant, and injected CTLA4Ig+anti-CD154 applied; (3)normal mice injected by corresponding isotype control IgG of CTLA4Ig and anti-CD154; (4)normal blank control mice. Each group had 15 mice. On day 0, mice of each group were irradiated lethally 8 Gy by linear accelerator, and the bone marrow cells of C57BL/6 labeled by fluorescence staining were injected via the tail vein. The fluorescent cell level in peripheral blood and organ tissue at different time points were detected by flow cytometry (FCM) for homing assessment. Survival rates and hematopoietic reconstitution were also monitored and recorded. The results showed that application of CTLA4Ig anti-CD154 could promote implantation of allogeneic HSC in sensitized recipients, induce the immune tolerance, prolong their survival time and accelerate the hematopoietic reconstitution within 28 days, compared with the sensitized group. It is concluded that applying CTLA4Ig and anti-CD154 can enhance the engraftment of HSCT and induce immune tolerance in the sensitized recipient after allogeneic HSCT and accelerate the hematopoietic reconstitution.
Abatacept ; Animals ; B7 Antigens ; antagonists & inhibitors ; CD28 Antigens ; antagonists & inhibitors ; CD40 Antigens ; antagonists & inhibitors ; CD40 Ligand ; antagonists & inhibitors ; Graft Rejection ; prevention & control ; Hematopoietic Stem Cell Transplantation ; Immune Tolerance ; Immunoconjugates ; pharmacology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Transplantation, Homologous

Objective: To investigate surgical strategies and the corresponding benefits for patients with perihilar cholangiocarcinoma(pCCA). Methods: A total of 81 patients with pCCA who underwent radical excision in the Department of Biliary and Pancreatic Surgery of Sun Yat-Sen Memorial Hospital between January 2014 and December 2021 were retrospectively collected.The cohort consisted of 50 male and 31 female patients,with an age of (62.5±11.5)years(range:26 to 83 years).Seventy-five cases were diagnosed with jaundice,60 of whom received preoperative biliary drainage,while 20 patients received portal vein embolization.Their serum bilirubin level within one week before the operation(M(IQR)) was 44.3 (41.9) μmol/L(range:8.0 to 344.2 μmol/L).Preoperative imaging examinations were performed to evaluate the Bismuth-Corlette type of pCCA,showing 3,6,21,27,and 24 cases of Bismuth-Corlette type Ⅰ,Ⅱ,Ⅲa,Ⅲb,and Ⅳ,respectively.The primary outcome was overall survival (OS),and the secondary outcomes were relapse-free survival (RFS),90-day postoperative morbidity and 90-day postoperative mortality.OS and RFS were estimated using the Kaplan-Meier method and compared by the Log-rank test.Significant prognostic factors were determined using univariate and multivariable Cox proportional hazard regression analyses. Results: In the cohort of 81 pCCA patients,67 cases(82.7%) underwent major hepatectomy while 3 cases received major hepatectomy combined with pancreaticoduodenectomy.Thirty-four patients underwent hepatectomy combined with vascular resection and reconstruction(18 cases of portal vein resection and reconstruction alone;9 cases of hepatic artery resection and reconstruction alone;7 cases of combination of portal vein and hepatic artery resection and reconstruction).Margin negative(R0 excision) were achieved in 53.1%(43/81) of these patients.The operation duration was (627±136)minutes(range:565 to 940 minutes),and the intraoperative blood loss was 400(455)ml(range:200 to 2 800 ml).The 90-day postoperative mortality was 3.7%(3/81).Grade 3-4 postoperative morbidity was 23.4% (19/81) according to the Clavien-Dindo classification of surgical complications.Up to the last follow-up at September 2022,the follow-up time was 34.0(24.2)months (range:0.4 to 103.6 months).Three patients who died within 90 days after surgery were excluded from the survival analysis.The median OS was 36.10 months (95%CI:18.23 to 42.97 months) and the 1-,3-and 5-year OS rates were 85.3%,46.8% and 27.3%,respectively.The median OS of 41 patients with negative margins was 47.83 months(95%CI:36.90 to 58.80 months) and that of 37 patients with positive margins was 20.47 months(95%CI:10.52 to 30.58 months).The median RFS of 70 patients with R0 and R1 resection was 24.50 months(95%CI:12.15 to 31.85 months)and the 1-,3-and 5-year RFS rates were 65.2%,45.7% and 29.9%,respectively.The median RFS of 41 patients with R0 resection was 38.57 months(95%CI:21.50 to 55.63 months) and that of 29 patients with R1 resection was 10.83 months(95%CI:2.82 to 19.86 months). Conclusions: The primary therapy for pCCA is radical surgical resection.A precise preoperative evaluation and sufficient preparation can reduce postoperative morbidity.Surgical treatment can achieve a better survival outcome by increasing the radical resection rate.