Objective To research the role of hypothalamocerebellar histaminergic projections in somatic‐visceral integration of rats .Methods A total of 120 SD rats were randomly divided to 6 groups :group A - F ;which separately microinjected saline (group A) ,triprolidine(selective antagonist of H1 receptor ,group B) ,2‐PyEA(selective agonist of H1 receptor ,group C) ,ranitidine (selective antagonist of H2 receptor ,group D) ,dimaprit (selective agonist of H2 receptor ,group E) and histamine (group F) to the deep nuclei of rats .The abilities of rats somatic‐visceral integration were observed when hypothalamocerebellar histaminergic pro‐jections were obstructed or enhanced compared to group A .Results The somatic‐visceral integration of rats were obstructed obvi‐ously in group D and enhanced in group E & group F compared to group A ,while group B and group C had no differences compared to group A .Conclusion Hypothalamocerebellar histaminergic projections play a key role in somatic‐visceral integration through the way of H2 receptors .

Objective To evaluate the diagnosis and therapy of intestinal obstruction caused by multiple jejunal diverticula.Methods Clinical data of intestinal obstruction caused by multiple jejunal diverticula in 18 cases admitted from Jan 2001 to Aug 2012 were analyzed retrospectively.Results All patients experienced the symptoms of abdominal pain and abdominal distension,5 cases experienced nausea and vomit,2 cases had a fever.Plain abdominal X-ray series demonstrated distension of small bowel and multiple air-fluid levels,5 cases with anemia,the average value of hemoglobin was (9.5 ± 1.0) g/L,B-ultrasonography revealed bowel dilatation in 7 out of 15 cases,9 cases underwent abdominal computed tomography and all had positive sign of small bowel distension and multiple air-fluid levels,mesenteric volvulus was suspected in 1 case.All patients underwent laparotomy,the diagnosis of multiple jejunal diverticula were confirmed clinically and by the pathology.In a follow-up ranging from 2 to 25 months,1 case died,the others were symptoms free.Conclusions Multiple jejunal diverticula was a less common disorder,occurring mainly in old man,with a low preoperative definite diagnosis.Resection of affected intestinal segment with primary anastomosis results in satisfactory prognosis.

Objective To investigate the feasibility and safety of autologous tumor tissue lysate loading den-dritic cells(DC) for the treatment of hepatocellular carcinoma (HCC). Methods The monocytes-derived DC were induced and antigen loaded with tumor tissue lysate to produce DC vaccine. Vaccination and clinical observation were conducted in 12 HCC patients for 41 times. Results The average output of DC was 1.69×107(1.69×107±9.44×106>) from 90 ml peripheral blood. 63.41% (26/41)patients appeared to develop delayed-type hypersensitivity after intradermal injection. After an average of 9 months follow up, 1 patient out of 4 recurrence and metastasis pa- tients survived for 17 months. The other three patients progressed. Out of 8 patients undergoing immunotherapy post- operatively,6 patients had no signs of recurrence and the others were found to have liver rceurrence and progression. Conclusion DC based immunotherapy is safe and feasible,with no side effects,which can be applied in the immu- notherapy strategy of HCC patients.

Objective:To investigate the most effective strategy for mature induction of dendritic cells.Methods:Human monocyte-derived dendritic cells were induced in the presence of cytokines GM-CSF and IL-4. On day 6, the immature DCs were pulsed with each of CD40L, LPS, TNF-? or a cocktail of cytokines(TNF-?, IL-6, IL-1?, PGE2). DCs were harvested after 24 h induction. The surface markers for maturation CD80,CD83,CD86 and HLA-DR were detected by FCM. FITC-Dextra endocytic activity was measured by FCM. IL-12 production was detected by ELISA. The capacity of DCs for T cell activation was detected by MTT assay.Results:CD40L,LPS,TNF-? and the cocktail of cytokines all could induce DCs’ maturation. The most effective scheme for induction of maturation was the cocktail of cytokines, and the expression rate of CD83 was up to 66.91%(P

Objective To evaluate the therapeutic effect of narrow band-ultraviolet B (NB-UVB) alone or in combination with interferon-alpha-2b (INF-alpha-2b) for mycosis fungoides (MF),and to assess the correlation between the therapeutic effect and peripheral blood regulatory T (Treg)/T helper type 17 (Th 17) cells.Methods Thirty-three patients with stage ⅠA to ⅡA MF were randomly divided into two groups:NB-UVB group (n =15) receiving NB-UVB radiation alone,combined group (n =18) treated with NB-UVB radiation and intramuscular injection of INF-alpha-2b.Ten healthy volunteers were selected as the control group.Peripheral blood samples were collected before and 9 months after the start of treatment.Flow cytometry was performed to determine the percentages of Treg cells and Th17 cells.Statistical analysis was carried out by using t test,one-way analysis of variance,and Fisher's exact test.Results The average treatment duration was 9 months among these patients.Therapeutic outcomes were significantly better in the combined group than in the NB-UVB group (P =0.023).Among the 15 patients in the NB-UVB group,6 achieved complete remission,3 partial remission,6 showed no response;of the 18 patients in the combined group,12 experienced complete remission,5 partial remission,and 1 showed no response.Before the treatment,the percentages of both Treg and Th17 cells in peripheral blood were significantly higher in the NB-UVB group and combined group than in the control group (both P ＜ 0.05),but similar between the NB-UVB group and combined group (both P ＞ 0.05).After the treatment,the percentages of both Treg and Th17 cells in the NB-UVB group and combined group significantly decreased compared with those before the treatment,but were still higher than those in the control group (both P ＜ 0.05).Additionally,the degree of decrease in the percentages of Treg and Th17 cells was significantly greater in the combined group than in the NB-UVB group (both P＜ 0.05).The seven patients with no response also showed a significant decrease in the percentage of Treg cells (P ＜ 0.05),but no obvious changes in that of Th 17 cells (P ＞ 0.05) after the treatment.Conclusions The therapeutic effect of NB-UVB radiation combined with intramuscular INF-alpha-2b is superior to that of NB-UVB radiation alone for MF,which may be associated with the degree of decrease in peripheral blood Treg and Th 17 cells.

Objective To measure the expression of interleukin-13 (IL-13) and its receptors in mycosis fungoides (MF) lesions,and to investigate their clinical significance.Methods A total of 34 paraffin-embedded specimens of MF,which was confirmed by clinical and histopathological features,immunophenotyping and/or T-cell receptor gene rearrangements,were collected from Hangzhou Third People's Hospital between January 2010 and March 2016.According to the tumor-node-metastasis (TNM) staging system,5 patients were at stage I A,9 at stage Ⅰ B,17 at stage Ⅱ A,and 3 at stage Ⅱ B.Ten normal skin tissue specimens served as controls.Immunohistochemical study was conducted to measure the expression of IL-13,IL-13Rα1 and IL-13Rα2.Results IL-13,IL-13Rα1 and IL-13Rα2 were all expressed in atypical lymphoid cells and epidermotropic lymphoid cells in MF lesions at various stages.IL-13Rα2 was highly expressed in all the MF lesions.None of IL-13 and its receptors were expressed in normal skin tissues and lymphocytes.The expression rates of IL-13 and its receptors in MF lesions increased along with the progression of MF.Additionally,the expression rates of IL-13 (10.00％ ± 3.14％),IL-13Rα1 (21.43％ ± 6.88％) and IL-13Ro2 (31.14％ ± 6.38％) significantly decreased in MF lesions at stage Ⅰ compared with those at stage Ⅱ (27.50％ ± 11.00％,39.45％ ± 9.43％,44.40％ ± 11.15％,respectively,all P ＜ 0.05),but no significant differences were observed between stage Ⅰ A and Ⅰ B,or between stage Ⅱ A and Ⅱ B (P ＞ 0.05).Conclusion IL-13 and its receptors,especially IL-13Rα2,may be expected to serve as biomarkers for early diagnosis of MF and prediction of its biological behaviors.

Objective To investigate the changes of T helper type 17 (Thl7) cells and regulatory T (Treg) cells in different stages of mycosis fungoides.Methods Flow cytometry was performed to determine the percentage of Treg and Th17 cells in peripheral blood from 28 patients with mycosis fungoides (MF),13 patients with large plaque parapsoriasis (PP),17 patients with lichen planus (LP) and 10 healthy human controls,and immunohistochemistry to detect the expressions of forkhead box protein 3 (FOXP3) and interleukin (IL)-17 in tissue specimens from 40 patients with MF,13 with PP,17 with LP and 10 healthy human controls.Statistical analysis was carried out by one-way analysis of variance and Pearson correlation analysis.Results As far as the percentage of Treg cells in peripheral blood was concerned,MF,PP and LP patients were significantly higher than the healthy controls (8.09％ ± 1.68％,6.53％ ± 1.67％ and 2.84 ％ ± 1.16％ vs.1.01％ ± 0.35,all P＜ 0.05),PP patients were higher than LP patients and healthy controls (both P ＜ 0.05),and LP patients were higher than healthy controls (P ＜ 0.05).The percentage of Th17 cells in peripheral blood was significantly increased in MF patients compared with PP patients,LP patients and healthy controls (3.22％ ± 0.82％ vs.2.46％ ± 0.79％,1.38％ ± 0.47％ and 0.59％ ± 0.30％,all P ＜ 0.05).Elevated expression rate of FOXP3 was observed in MF,PP and LP lesions as compared with normal skin (14.94％ ± 4.46％,11.95％ ± 4.72％,6.32％ ± 2.81％ vs.3.43％ ± 1.79％,all P ＜ 0.05),and in MF and PP lesions compared with LP lesions (both P ＜ 0.05),but no significant difference was observed between MF and PP lesions (P ＞ 0.05).There was a significant increase in the expression rate of IL-17 in MF lesions compared with PP lesions,LP lesions and normal skin (15.89％ ± 4.27％ vs.12.02％ ± 3.34％,4.84％ ± 1.93％ and 2.62％ ± 0.89％,all P ＜ 0.05).The Th17/Treg ratio in peripheral blood was significantly lower in MF and PP patients than in LP patients and healthy controls (0.41 ± 0.11 and 0.39 ± 0.12 vs.0.50 ± 0.06 and 0.57 ± 0.19,all P ＜ 0.05).A positive correlation was observed between the proportion of Thl7 cells and Treg cells (r =0.423,P ＜ 0.05) in patients with early-stage MF,but not in those with tumor-stage MF.The proportion of Th17 cells decreased,but that of Treg cells continuously increased in patients with tumor-stage MF.However,no significant difference was noted in the proportion of Thl7 cells or Treg cells among patients with different stages of MF.Conclusion The imbalance between Treg and Th17 cells may be involved in the occurrence and development of MF.

Objective To explore the effect of histamine on neurons in the hippocampal C1 area of SD rats and the behavior of depressive SD rats. Methods The effect of histamine on the discharge frequency of neurons in hippocampal C1 area was observed by in vitro extracellular recording of brain slices. The effect of endogenous histamine released to hippocampal neurons was observed through in vivo extracellular recording after the hypothalamus was electrically stimulated. In addition, the changes of motor activity and the ability of spatial memory of the rats with depression after microinjection of histamine into the hippocampal C1 area were observed by open-field test and Morris water maze test. Results The results of in vitro extracellular recording of brain slices showed that the hippocampal neurons were excited by histamine via H1 receptors rather than H receptors in a concentration dependent manner. The results of in vivo extracellular recording showed that endogenous histamine had a bidirectional effect, which means a short-term excitatory effect followed by a long-term inhibitory effect on the hippocampal neurons. After injected with histamine in the hippocampal C1 area,

AIM: To investigate the immune stimulation capacity of B7-H1 blockade on immature dendritic cells (DCs) in vitro. METHODS: The human monocyte-derived dendritic cells were induced in the presence of cytokine GM-CSF and IL-4. The expression of B7-H1 was detected by FCM. On blockade of B7-H1, the maturation and endocytic activity, T cells stimulatory proliferation capacity, IL-12 production, T cell differentiation effect of DCs were detected by FCM, MTT assay, ELISA and ELISPOT, respectively. RESULTS: The expression of B7-H1 was increased with the induction of DCs. On day 7, the positive expression was 54.12%, and the TNF-? induced mature DCs had the positive expression rate of 83.64%. The blockade of B7-H1 on immature DCs had sharply increased their T cells stimulatory proliferation capacity and IL-12 production, and efficiently induced the development of Th1/Tc1 cells, but had no effect on their maturation and endocytic activity. CONCLUSION: The blockade of B7-H1 on immature DCs increases its immune stimulation activity. It is valuable to investigate the antitumor immune responses of DCs vaccine with B7-H1 blockade.

Objective To explore the molecular mechanism of Xiaojin Pills in the treatment of breast cancer using an integrated network pharmacology and experimental verification.Methods The chemical components and potential targets of Xiaojin Pills were obtained from TCMSP,TCM-ID,ETCM and SwissTargetPrediction databases.Breast cancer related targets were collected from GeneCards,OMIM and KEGG databases.The overlapped targets were imported into STRING database to analysis a protein-protein interaction(PPI).The key targets of PPI networks were screened based on node topology parameter values through Cytoscape 3.8.0.DAVID database was used to analyze the GO and KEGG pathway enrichment to build drug-chemical components-key targets-signaling pathway network.The breast cancer cell lines MDA-MB-231 and SK-BR-3 were used to study the effects of Xiaojin Pills extract on cell apoptosis,migration and invasion,and to verify the key pathway obtained by enrichment analysis.Results Totally 181 chemical components in Xiaojin Pills were obtained,including quercetin,myricetin,pinocembrin and β-sitosterol.615 potential targets were identified for the anti-breast cancer effects of Xiaojin Pills.After overlapping,170 key targets against breast cancer were identified based on the topological analysis,which included SRC,ERK1/2,AKT1,EGFR,etc.KEGG analysis enriched pathways including pathways in cancer,MAPK signaling pathway,endocrine resistance,PI3K-AKT signaling pathway,EGFR tyrosine kinase inhibitor resistance,apoptosis,and HIF-1 signaling pathway,which may play important roles in the therapeutic effects of Xiaojin Pills against breast cancer.GO enrichment was involved in protein phosphorylation,inflammatory response,negative regulation of apoptosis,and positive regulation of ERK1 and ERK2 cascades.Cell experiments showed that Xiaojin Pills further induced mitochondria-dependent apoptosis by inhibiting the activation of MAPK and PI3K-AKT pathways.At the same time,the expressions of ZO-1 and β-catenin increased,and the epithelial-mesenchymal transformation process was reversed to inhibit the metastasis of breast cancer cells.Conclusion The key targets and signaling pathways of Xiaojin Pills in the treatment of breast cancer are studied through network pharmacology combined with in vitro experiments,which provided a basis for further study of its pharmacodynamic material basis,mechanism of action and clinical application.