Objective To investigate the effect of high mobility group box-1 protein (HMGB1) on inter-leukin-2 (1L-2) and interleukin-2 receptor α (IL-2α) expressions in human T lymphocytes and its potential regulat-ing mechanism in vitro. Method Human T lymphocytes were isolated and suspended, the cells were cultured with 20 μg/mL phytohemagglutinin (PHA) in 5% CO_2 at 37 ℃, recombinant human HMGB1 (rhHMGB1, 0, 10, 100, 1000 ng/mL) was added with the PHA and cultures were centrifuged at 12 and 48 hours for cell collect-ing. Reverse transcription polymerase chain reaction (RT-PCR) amplification was perfomed to determine gene ex-pressions of IL-2, IL-2Rα. IL-2, sIL-2R protein levels in cell culture supematants were measured by ELIZA. Re-sults After coincubated with rhHMGB1 (10, 100, and 1000 ng/mL) for 12 hours, IL-2 levels in cell culture su-pernatants respectively were 0 . 064 ± 0. 017 μg/L, 0.076±0.033 μg/L, and 0.061 ±0.02 μg/L, which were significantly higher compared with the untreated cells (0.045±0.011 μg/L, P < 0.05 or P < 0.01). Mean-while, IL-2 mRNA expression was markedly up-regulated following rhHMGB1 stimulation in various doses (F = 4.6872, P < 0.01). At 48 bourn, however, both IL-2 mRNA expression and protein production tended to de-crease along with an increased dose of dd-IMGB1 stimulationn. IL-2/sIL-2R ratio in 1000 ng/mL rhHMGB1 was markedly lower than that in 10 ng/ml rhHMGB1 (0.036±0.015 vs.0.055±0.017, P <0.05), together with down-regulation of IL-2Rα mRNA expression(P <0.01). Conclusions These data indieated that HMGBI could marked influence the IL-2/IL-2R expression in human T lymphocytes. With the increase in stimulating doses and prolongation of time, HMGBI might down-regulate T cell-mediated immune response of human lymphocytes.

Aim To investigate the inhibitory effects of urocortin2 (UCN2) on ventral tegmental area (VTA) nervous activity of morphine addiction rats and the mechanism. Methods Morphine addiction rats and the microiontophoresis method were used to observe the effects of UCN2 on VTA neuron′s spontaneous dis-charge changing rule, as well as the inhibitory effects of UCN2 on DA neuron′s cluster spontaneous dis-charge, to identify UCN2 and DA on the same VTA neuron. Morever, the inhibitor of corticotropin-regula-ting factor′s receptor ( CRF-2 R ) and the blocker of protein kinase A ( PKA ) , AST-2 B and H89 , were used to investigate the effects of UCN2 on VTA neuron′s of morphine addiction rats. Results UCN2 could inhibit the firing rate 82% (31/38) of the tested VTA neuron ( P<0. 01 ) , the discharge frequency changed from (20. 89 ± 2. 86) Hz to (13. 66 ± 3. 93) Hz (P<0. 01). Further, the inhibitor of PKA, AST-2B and H89 could ablolish the inhibitory effects of UCN2 . Morever, the excitatory firing of VTA neurons was at-tenuated by addition of UCN2 , while AST application could inhibit the UCN2′s inhibitory effects. Conclu-sion UCN2 could regulate the effects of VTA via PKA pathway and may thereby contribute to the improvement of drug addiction.

Objective To investigate the intervention of Chinese herb Shengjiang San(SJS)for the imbal-ances of serum Th1/Th2 cells and related regulatory factors in sepsis patients. Methods Fifty-five sepsis patients were randomly divided into two groups:conventional treatment group of 27 cases and SJS group of 28 cases. Nine cases of healthy volunteers were enrolled as control group in the study. Cases of conventional group were treated with western medicine only and cases of SJS group were treated with both western medicine and Chinese herb SJS (100 mL twice one day). The therapy course of both groups was 3 days. Score of Chinese medical syndromes ,se-rum level of leukocyte count,C reactive protein(CRP)and procalcitonin(PCT),serum T-bet,GATA-3,Th1&Th2 cells in the proportion of whole CD4+Th cells and Th1/Th2 ratio were compared respectively in each group be-fore and after treatment. Results There were significant differences between SJS group and conventional group in score of Chinese medical syndromes,serum level of leukocyte count,T-bet,GATA-3,Th1&Th2 cells in the pro-portion of whole CD4+Th cells and Th1/Th2 ratio(P<0.05 or P<0.01). There were no significant differences be-tween the two groups in serum level of CRP,PCT and GATA3. Compared with control group,there were signifi-cant differences in all indicators except GATA3 of the two groups(P<0.01). Conclusion Chinese herb Shengji-ang San has an effective benefits to Chinese medical syndromes,inflammatory reaction,the imbalance of serum Th1/T2 and related regulatory factors(T-bet&GATA-3)in sepsis patients.

Objective To investigate the effect of high mobility group box-1 protein (HMGB1) on cytokine expression in splenic dendritic cell (DCs). Method DCs isolated from the spleens of male Wistar rats were seed-ed on 96-well (1×10s cells/well) cell culture plates, and the cells were stimulated with HMGB1 for various length of time or in different concentrations. (1) The time-dependent response between HMGB1 and tumor necrosis factor-α(TNF-α) as well as interleukin-12 (IL-12) gene/protein expressions: 24 wells of DCs were dividedly into six groups including 24 h-normal controls (n=4), 48 h-normal controls (n=4), 72 h-normal controls (n=4), and 24 h-HMGB1 treated group (n=4), 48 h-HMGB1 treated group (n=4) as well as 72 h-HMGB1 treated group (n=4), respectively. Among three HMGB1-treated groups, DCs were stiraulated by 1 μg/mL HMGB1. DCs were denatured in cell culture plates to determine gene expression of IL-12 as well as TNF-α, and supernatants were harvested to determine Il-12 as well as TNF-α protein levels. (2) The dose-dependent response between HMGB1 and TNF-α as well as IL-12 gene/protein expressions: 16 wells of DCs were dividedly into four groups in-cludingnormal controls (n=4), 0.1 μg/mL HMGB1 treated group (n=4), 1 μg/mL HMGB1 treated group (n =4), and 10 μg/mL HMGB1 treated group (n=4), respectively. After stimulated for 48 h, DCs were dena-tured in cell culture plates to determine gene expression of IL-12 as well as TNF-α, and supernatants were harvest-ed to determine IL-12 as well as TNF-α protein levels. Total RNA was extracted from cells using the single-step technique of acid guanidinium thiocyanate-chloroform extraction according to the manufacturer' s instruction (Promega, Madison, WI). mRNA for TNF-α and IL-12 were quantified by SYBR Green two-step, real-time re-verse transeription-polymerase chain reaction taking glyceraldehyde-3-phesphate dehydrogenase (GAPDH) as an internal standard. Levels of IL-12 and TNF-α in cell culture supernatants were determined with ELISA, strictly fol-lowing the protocols provided by the manufacturer. Data were analyzed with a one-way ANOVA. A P-values <0.05 were considered statistically significant. Results After stimulation with 1 μg/mL HMGB1, IL-12 and TNF-α protein and gene expressions in rat splenic DCs were markedly up-regulated at 24 h to 72 h (P<0.05 or P<0.01), and the expression levels of IL-12 and TNF-α peaked at 48 h (P<0.01). When DCs were cultured in the presence of 0.1 μg/mL, 1 μg/mL, and 10 μg/ml HMGBI for 48 h, expressions of IL-12 and TNF-α were also significantly up-regulated (P<0.01), and values of these cytokines were highest in 1 μg/mL HMGB1-treated group (P<0.01). Conclusions These data suggest that HMGB1 appears to be a potential immunostimulatory signal that induced DC maturation, and HMGB1 stimulation can result in marked up-regulation of IL-12 as well as TNF-α synthesis and release in splenic DCs.

Objective:To investigate the clinical characteristics of renal injury caused by Guanxinsuhe pill(冠心苏合丸,including slander dutchanspipe root(青木香)).Methods:The clinical data of 27 patients with renal injury caused by Guanxinsuhe pill were studied.The clinical characteristics of the renal damage caused by Guanxinsuhe pill and the relationships between duration of illness,duration of taking Guanxinsuhe pill,anemia,renal atrophy and the progression of renal injury respectively were also analyzed.Results:Twenty-seven patients all taken Guanxinsuhe pill for suspicious coronary heart disease(CHD) and were diagnosed as chronic tubulointerstitial nephropathy accompanied with renal tubular acidosis,renal glucosuria and hyposthenuria after admission.Anemia was of more severity than renal dysfunction.Atrophic kidney occurred in 26 patients.The main complains were nonspecific symptoms including fatigue,anorexia,nausea,vomiting,etc.and accompanied with different degrees of chronic renal dysfunction.Twentyfour hours urinary protein excretions were 120-900 mg((490?250) mg).There were few red and white cells in uroscopy.Sixteen patients discontinuously took the Guanxinsuhe pill(one capsule,2-3 times a day) according to the pharmacopoeia for a long period.Ten patients took a dosage(two capsules,2-3 times a day) higher than that limited by the pharmacopoeia.Twenty-six patients took Guanxinsuhe pill discontinuously.One patient continuously took Guanxinsuhe pill until got renal dysfunction.The duration of taking Guanxinsuhe pill was 1-120 months(37.38?31.58),and the amount of 50-43 800 pills(4 576.00?8 830.54) were used,but the amount of pills of 3 cases was uncertain.The renal injury occurred within 30-216 months(95.41?56.74) after initiation of the drug taken.The renal injury obviously correlated with the anemia and the duration of taking Guanxinsuhe pill.Conclusion:Taking Guanxinsuhe pill can cause renal injury.The Guanxinsuhe pill dose indicated in pharmacopoeia also causes renal damage.The onset of renal injury is insidious.Most of the patients have the history of CHD and when the chronic nephropathy in most of such patients is detected,it is advanced to stage 5.The renal injury induced by Guanxinsuhe pill usually presents as chronic tubulointerstitial nephropathy,renal tubular dysfunction and anemia.Anemia is of more severity than renal dysfunction.The 24 hours urinary protein excretions of all the patients are lower than 1 000 mg daily.It is suggested to avoid taking Guanxinsuhe pill,adjust the prescription of Guanxinsuhe pill and do not use slander dutchanspipe root as an ingredient.

Objective To observe the expression law of cytokine signaling suppressors (SOCSs) mRNA in burn rats with Staphylococcus aureus sepsis and investigate their potential role in the pathogenesis of postburn sepsis. Methods Wistar rats were inflicted with 20% TBSA Ⅲ? scald, followed by Staphylococcus aureus challenge. Then, the expressions of SOCS1, SOCS2 and SOCS3 mRNA and interferon-? (IFN-?) levels in the liver and lungs were determined. Results With Staphylococcus aureus challenge after burn, IFN-? levels in the liver and lungs were significantly elevated and reached peak at the 0.5th and 6th hours, respectively (P

Objective To analyze the mutation status of K‐ras gene in colorectal cancer patients ,further more ,to provide guid‐ance for personalized therapy for colorectal cancer .Methods Nested and COLD‐PCR were used to detect the K‐ras mutations in 560 patients with colorectal cancer .Results In 560 colorectal cancer patients ,the total positive rate of K‐ras gene mutations was 27 .08% ,the mutation rate was 0 in 128 plasma samples and it was 27 .08% in 432 tissue samples .The mutate sites were G12S , G12C ,G12D ,G12A ,G12V ,G13R ,G13C ,G13D ,Q61K ,Q61L ,there were significant differences existed in different samples (P <0 .000 1) ;the mutation rate of 362 male patients was 20 .44% and the types of mutation include G12S ,G12C ,G12D ,G12V ,G13R , G13C ,G13D ,Q61K and Q61L .The mutation frequency was 21 .72% in 198 female patients ,the mutation points were G12S ,G12C , G12D ,G12A ,G12V ,G13R and G13D .There were no significant difference between different sex (P= 0 .722 7) ;the mutation fre‐quency was 20% in 80 youth patients including G12S ,G12C ,G12D ,G12V ,G13D and the mutation rate was 33 .07% in 127 middle age patients ,the points of mutation were G12S ,G12D ,G12A ,G12V ,G13R ,G13C ,G13D ,Q61K ,Q61L ,the mutation frequency was 16 .71% in 353 old age patients ,the types of mutation include G12C ,G12D ,G12V ,G13R ,G13D ,the difference was significant a‐mong different age patients (P= 0 .000 5) .Conclusion The total rate of mutations is 27 .08% in 560 colorectal cancer patients ,and the main points of mutation is G12D ,G12V ,G13D .There are significant differences in different type of samples as well as in differ‐ent ages ,but no statistical significance in different sex patients .

Professor HE Tianyou's unique understanding and treatment characteristics for intractahle facial paralysis are introduced. In clinical practice professor HE highly values acupoint selection and manipulation application, and integrates Chinese and western medicine to flexibly choose acupoints and formulate prescriptions according to syndrome differentiation and location differentiation, besides, he creates several specialized manipulation methods including "tug-of war opposite acupuncture method" and "tractive flash cupping". Based on strengthening body and dredging collaterals. more attention is given on stimulation to local paralyzed facial nerves; meanwhile acupuncture and medication are combined to improve clinical efficacy. During the treatment, the important role of psychological counseling on patient's anxiety is emphasized, and comprehensive treatment is given physically and psychologically in order to achieve the purpose of total rehabilitation.
Acupuncture Points ; Acupuncture Therapy ; Adult ; Drugs, Chinese Herbal ; administration & dosage ; Facial Paralysis ; drug therapy ; therapy ; Female ; Humans ; Medicine

Aim To investigate the effects of cholecystokinin-octopeptide on IL-12 secreted in murine bone marrow-derived dendritic cells induced by lipopolysaccharide.Methods The CCK receptor subtypes were investigated by immunofluorescence in murine bone marrow-derived dendritic cells.Enzyme linked immunosorbent assay and Western blot were used to estimate the contents of IL-12 and p38MAPK activity.Results CCK-1R and CCK-2R were detected in BM-DC;CCK-8(at concentrations of 10-10,10-8,10-6 mol?L-1)could significantly increase the secretion of IL-12 in the LPS-induced DC, and LPS-activated p38MAPK activity in a dose-dependent manner.The effect of CCK-8 was reduced partially by CR1409(a CCK-1R antagonist) and CR2945(a CCK-2R antagonist).Conclusion CCK-8 could dose-dependently increase the expressions of IL-12 in LPS-induced DC,probably by promoting p38MAPK activity and the effect was mediated by CCK1 and CCK2 receptors.

AIM: To study the effects of cholecystokinin octapeptide (CCK-8) on T-lymphocyte subsets in keyhole limpet haemocyanin (KLH)-immunized mice.METHODS: Female BALB/c mice were immunized with KLH and injected with different dosages of CCK-8 or saline simultaneously. Positive CD4+and CD8+ T cells in peripheral blood or splenocytes were measured by flow cytometry. The production and mRNA level of Th1 cytokine, IFN-? and Th2 cytokine, IL-4 in the splenocytes were evaluated by ELISA and RT-PCR, respectively. In addition, lung tissue sections were made with HE staining.RESULTS: CCK-8 downregulated the positive CD4+and CD8+ T lymphocytes both in peripheral blood and in splenocytes in KLH-immunized mice, resulting in the reduction of CD4+/CD8+ ratio.CCK-8 improved the production of IFN-?, elevated IFN-? gene transcription, whereas cut down the production of IL-4, decreased IL-4 gene transcription. CCK-8 lightened the pulmonary inflammation induced by KLH.CONCLUSION: CCK-8 inhibits CD4+T lymphocytes activation, Th2 cytokine mRNA expression and protein production in KLH-immunized mice, indicating that CCK-8 modulates adaptive immune response.