ObJective To locate the cysteine-rich domains(CRD) of murine 4-1BB binding to its natural ligand. Methods A serial soluble extracellular CRDs of routine 4-1BB and 4-1BBL fusion proteins was constructed and prepared. The binding of purified 4-1BB-Igs to 4-1BBL and 4-1BB monoclonal antibody were tested using ELISA assay and Western blot analysis. Blocking experiment with 4-1BBL and 4-1BB mon-oclonal antibody was performed by ELISA assay. Results All truncated overlapped proteins containing ex-tracellular CRD Ⅱ of murine 4-1BB were able to bind to 4-1BBL by ELISA assay, excepting the CRD Ⅰ do-main alone. A 4-1BB monoclonal antibody proved to block the interaction of 4-1BB and 4-1BBI, was also able to bind to CRD Ⅱ. Conclusion Murine 4-1BBL whose specificity was mapped to CRD Ⅱ of 4-1BB ex-tracellular region with a possible conformational structure.

Objective To evaluate the safety and efficacy of radical antithrombotic therapy(aspirin,clopidogrel and tirofiban) in the treatment of senile (≥ 75 years) non-ST-elevation acute coronary syndrome (NSTEACS).Methods A total of 146 senile patients with NSTEACS were divided into observation group (70 cases) and control group(76 cases) by random digits table method.The control group was given aspirin,low molecular weight heparin and clopidogrel,and the observation group was added tirofiban (intravenous loading dosage:0.4 μ g/ (kg· min) for 30 min and then maintaining 0.1 μ g/ (kg· min) for 48-72 h).The occurrence of major adverse cardiovascular events (MACE) within 30 d after treatment,7 d angina pectoris control rate and adverse reactions were observed in two groups.Results The occurrence rate of MACE within 30 d after treatment was 5.7％ (4/70) in observation group,and 14.5％ (11/76) in control group,and there was significant difference (P ＜ 0.05).The 7 d angina pectoris control rate was 92.9％ (65/70) and 76.3 ％ (58/76) in observation group and control group,and there was significant difference (P ＜ 0.05).The incidence of bleeding was 10.0 ％ (7/70) and 7.9 ％ (6/76) in observation group and control group,and there was no significant difference (P ＞ 0.05).There were no major bleeding events in two groups.One case had thrombocytopenia in observation group,but there was no significant difference compared with that in control group(P ＞ 0.05).Conclusion Triofiban on the basis of aspirin and clopidogrel in the treatment of senile NSTEACS is effective and safe.

Objective To analyze the allergens and sex ,age distribution of chronic urticaria in Chengdu .Methods Totally 859 patients with chronic urticaria were tested with 13 kinds inhaled allergens and 15 kinds of food allergens by skin prick test .Re‐sults The top 5 allergens were:dermatophagoides farinae、dermatophagoides pteronyssinus ,cockroach ,shrimp and sea‐crab .The positive rate of dermatophagoides farinae was 58 .7% ,dermatophagoides pteronyssinus which was 55 .1% took second place .No difference was found between sex ,more inhaled allergens were found positive than food allergens in both sex groups .The positive rate was higher in people younger than 60 .Conclusion Dermatophagoides farinae ,dermatophagoides pteronyssinus ,cockroach , shrimp and sea‐crab are the commonest allergens in Chengdu .The skin prick test is important in the individualized treatment of chronic urticaria and health education .It may also be helpful in the management of chronic allergic skin diseases .

Objective To investigate the effects of penehyclidine hydrochloric(PHC)pretreatment on the expression of β-arrestin-2 in the lung tissue in sepsis-induced acute lung injury in mice.Methods Thirty female Ktmming mice,aged 6 weeks,weighing 18-20 g,were randomly divided into 3 groups(n =10 each):sham operation group(group S); sepsis group(group CLP)and penehyclidine hydrochloric pretreatment group(group PHC).Sepsis was induced by cecal ligation and puncture(CLP)in groups CLP and PHC.Penehyclidine hydrochloric 0.45 mg/kg was injected intraperitoneally at 1 h before CLP in group PHC.While the equal volume of normal saline was given instead of penehyclidine hydrochloric in groups S and CLP.At 12 h of CLP,the animals were sacrificed,and the lung tissues were removed for determination of MPO activity(by colorimetry),IL-6 content(by ELISA),β-arrestin-2 mRNA and protein expression(by RT-PCR and Western blot respectively).Blood samples and bronchoalveolar lavage fluid were collected to calculate pulmonary vascular permeability index(PV PI).Results Compared with group S,PVPI,IL-6 content and MPO activity were significantly increased,the expression of β-arrcstin-2 protein was significantly down-regulaled while the expression of β-arrestin-2 mRNA was up-regulated in group CLP,and PVPI,IL-6 content and MPO activity were significantly incrcased,the expression of β-arrestin-2 protein was significantly up-regulated,while the expression of β-arrestin-2 mRNA was down-regulated in group PHC(P ＜ 0.05).Compared with group CLP,PVPI,IL-6 content,and MPO activity were significantly decreased,the expression of β-arrestin-2 protein was significantly up-regulated,while the expression of β-arrestin-2 mRNA was dow n-regulated in group PHC(P ＜ 0.05).Conclusion PHC pretreatment can attenuate the lung injury induced by sepsis in mice through up-regulating the expression of β-arrestin-2 protein.

Objective To investigate the growth inhibition and radiosensitization of Celecoxib in hu-man nasopharyngeal carcinoma cell line CNE-2. Methods CNE-2 growth inhibition by Celecoxib was eval-uated by MTT method. Apoptosis-related changes in morphology were observed by transmission electron mi-croscopy (TEM). Cell cycle distribution and apoptosis rate were measured by flowcytometry (FCM). The ex-pression of COX-2 protein was observed by SP method after the treatment of Celecoxib. Cells were randomly planted into four groups: irradiation control(Ci), drug group(Cd), irradiation group(R), and Celecoxib plus irradiation group(D+R). Single irradiation of 2,4,6,8,and 10 Gy were administered for colonogenic assay. Cell cycle distribution and apoptosis rate were analyzed at 6 Gy irradiation. Results The growth of CNE-2 cell was inhibited by celecoxib in a dose-and time-dependent manner, the IC50 was 80 μmol/L After the treatment, cell ratio of GO and G, phases was increased (47.03±2.76 vs 56.17±1.95, t=4.68, P= 0.010), whereas the ratio of S and G2/M phases was decreased (33.07±1.86 vs 24.87±1.76, t=5.54, P = 0.010; 19.30±0.53: 17.73±0.83, t=2.75, P=0.050), and the apoptosis rate was increased (1.57±0.47:10.47±0.31, t = 27.39, P = 0.000) in a dose-dependent manner. Apoptosis with nuclear chromatin condensation, fragmentation and cell shrinkage was found by TEM. SP method showed that Celeib decreased COX-2 expression (17.48±0.34 vs 12.82±0.51,t=13.20,P =0.00). The sensitivity ratio(D0) was 1.15. FCM showed that the percentage of cells in G2/M phase was significanty more in R and D+R groups than in Ci and Cd groups (68.00±1.65,54.27±5.74,17.60±0.80,14.86±1.23, t=47.70,P=0.000; t=11.63, P=0.000), and also significantly different between R group and D + R group (t=3.99, P= 0.020). The apoptosis rate was higher in R and D + R groups than Ci and Cd groups(4.83±0.97,9.50± 1.35,1.33±0.86 and 2.28±0.42,t=4.67,P=0.010;t=8.81, P=0.000), D + R group than R group(t =4.85,P=0.010). Conclusions Celecoxib can markedly inhibit the growth and induce apoptosis in CNE-2 cells,which may depend on COX-2 pathway. Celeeoxib potently enhances the radiosensitivity of CNE-2 cells,which may due to the repair inhibit of radiation-induced DNA damage, inhibit of cell proliferation,and enhancement of cell apoptosis after irradiation.

Objective To investigate the characteristics of traditional Chinese medical syndromes and syndrome patterns in diabetic nephropathy (DN) from delta regions of pearl river. Methods A cross-section and retrospective trial was carried out in 292 DN inpatients from four hospitals(the First Affiliated Hospital of Guangzhou University of TCM, Foshan Hospital of TCM, Shenzhen Hospital of TCM, and Zhongshan Hospital of TCM) from January, 2003 to December, 2005. Demographic information, past history, symptoms and laboratory parameters of the patients were collected.Results Of the enrolled 292 patients, there were 14 in the clinical stage Ⅱ, 42 in stage Ⅲ, 210 in stage Ⅳ, and 26 in stage Ⅴ. The manifestations of TCM syndrome were dominated as lassitude and weakness (69.9%), loss of energy (54.1%), dry mouth (49.3%), aching waist (42%), dry mouth and throat (38%), numbness of extremities (69.9%), dry eyes (33.2%) and dry stool (31.5%), but the signs of frequent sighing, night sweating and tidal fever were less found. For the deficiency syndrome patterns, there existed deficiency of kidney (37.7%), spleen (27.7%), heart (8.9%), liver (3.8%) and lung (2.1%) when differentiating the syndromes according the five zang-organs; there was deficiency of yin (32.5%), Qi (19.5%), blood (9.6%) and yang (8.2%) when differentiating the syndromes according to Qi, blood, yin and yang; there were complex deficiency syndromes such as deficiency of Qi and yin (40.4%), deficiency of spleen and kidney Qi (26.4%), deficiency of lung and kidney Qi(4.8%) and deficiency of yin and yang (3.8%). For the excess syndrome patterns, there were blood stasis syndrome (49.7%), damp-heat syndrome (33.2%), water-dampness syndrome (25%), and Qi stagnation syndrome (7.5%). Conclusion The syndrome patterns of DN present the following characteristics: leading deficiency syndromes being kidney deficiency and spleen deficiency when differentiating the syndromes according the five zang-organs, being yin deficiency and Qi deficiency when differentiating the syndromes according to Qi, blood, yin and yang, and complex deficiency syndromes being Qi-yin deficiency and spleen-kidney Qi deficiency. The leading excess syndromes are blood stasis syndrome and damp-heat syndrome. The manifestations of DN are characterized by the concurrence of deficiency and excess, cold and heat. The pathogenesis of DN is classified into deficiency in the root and excess in the branch. The deficiency syndrome mainly involves Qi and yin, and is located in spleen and kidney. The excess syndrome is characterized by blood stasis and damp-heat, and by water-dampness and dampness-turbidity in the late stage of DN. In one word, the pathogenesis of DN is characterized by deficiency of Qi and yin, and blood-stasis blocking kidney collaterals.

Objective To analyze and investigate the diagnosis method and treatment measures of insulinoma.Methods The clinical data in 16 cases of insulinoma in the hepatobiliary surgery of our institute from Dec.2008 to Dec.2015 were retrospectively analyzed.Results All cases had typical Whipple′s triad,the biochemical examination suggested that all cases had plasma glucose decrease and plasma insulin increase during the onset period.The imaging examinations included preoperative ultrasonography,enhanced CT,MRI,EUS and intraoperative ultrasound (IOUS).Their diagnosis positive rates were 56.3% (9/16),77.8% (7/9),80.0% (8/10),100.0%(2/2) and 100.0%(5/5) respectively.Sixteen cases all received surgical treatment,including local tumor excision in 6 cases,simple distal pancreatectomy in 3 cases,distal pancreatectomy plus splenectomy in 5 cases and pancreaticoduodenectomy(PD) in 2 cases.Plasma glucose level in all cases was significantly increased after tumor removal.Postoperative follow-up lasted for 1-5 years without recurrence and metastasis.Conclusion The first choice of insulinoma localization diagnosis is CT and MRI,intraoperative palpation combined with IOUS is the main method of intraoperative localization,and the surgical resection is the only effective method for treating this disease.

Objective To evaluate the effect of electro-acupunctare at Zusanli on liver injury in a rat model of sepsis after scald.Methods Fifty SPF male Sprague-Dawley rats,weighing 200-250 g,aged 2-3 months,were randomly divided into 5 groups (n =10 each) using a random number table:control group (group C); sepsis after scald group (group SS); electro-acupuncture at Zusanli group (group E); electric stimulation of non-acupoint group (group NE); electro-acupuncture at Zusanli + α-bungarotoxin (α-BGT) group (group α-BGT).The rats were subjected to a third degree scald covering 20％ of the total body surface area (TBSA) and sepsis was induced with muramyl dipeptide (MDP) 5 mg/kg injected into the femoral vein immediately after scald.Electro-stimulation (3 V,2 ms,3 Hz) of bilateral Zusanli acupoints or non-acupoints was performed for 12 min every 8 h for 2 consecutive days starting from the time point immediately after MDP was injected in group E.In group NE,electro-stimulation was performed at the points 5 mm lateral to the acupoints of bilateral Zusanli and the method was similar to those previously described in group E.In group α-BGT,α-BGT 1.0 μg/kg (in normal saline 1 ml) was injected into the femoral vein before electric stimulation,then electro-stimulation was performed and the method was similar to those previously described in group E.After 48 h of continuous stimulation,liver specimens were obtained for microscopic examination of the pathological changes.The blood samples were obtained from the abdominal aorta for determination of the levels of serum ALT and AST,tumor necrosis factor-alpha (TNF-α),and high mobility group box-1 (HMGB1) (by ELISA),and expression of Nod-like receptor 2 (NLR2) mRNA (by RTPCR) and receptor-interacting protein 2 (RIP2) in the liver tissues (by Western blot).Results The serum ALT and AST,TNF-α and HMGB-1 levels and NLR2 mRNA and RIP2 expression in liver tissues were significantly higher in SS group than in C group.Compared with SS group,the serum ALT,AST,TNF-α and HMGB1 levels were significantly decreased and NLR2 mRNA and RIP2 expression was down-regulated in E group,and no significant changes were found in the parameters mentioned above in NE group.Compared with E group,the serum ALT,AST,TNF-α and HMGB1 levels were significantly increased,and NLR2 mRNA and RIP2 expression was up-regulated in α-BGT group.The pathological changes in liver tissues were significantly reduced in group EA as compared with SS,NE and α-BGT groups.Conclusion Electro-acupuncture at Zusanli can reduce liver injury in a rat model of sepsis after scald and inhibition of NLR2/RIP2 signaling pathway and activation of cholinergic antiinflammatory pathway mediated by α7 nicotinic acetylcholine receptors in liver tissues may be involved in the mechanism.

Objective To investigate the influence of total progressively motile sperm count(TPMSC) after treatment on clinical outcomes of intrauterine insemination(IUI) with the husband′s sperm in ovulation-promoting cycles.Methods The clinical data in 4179 cases undergoing IUI with the husband′s sperm in ovulation-promoting cycles were retrospectively analyzed.The correlation between clinical pregnancy rate and TPMSC was analyzed.Results Among all the clinical data,TPMSC was to 100×106 in occasional live sperm.TPMSC<0.15×106 was in 15 cases,1 case had pregnancy (live sperm was occasionally seen on IUI day after sperm processing).Ten cases of TPMSC >60×106 had no pregnancy.A total of 4 154 cases of TPMSC (0.15-60.00)×106 were analyzed.The female age,duration of infertility,number of follicles and endometrial thickness(EDM) had no statistical differences among various groups.The clinical pregnancy rate was 13.5%(576/4 154),the group with the highest clinical pregnancy rate was (5.00-<10.00)×106.But there was no statistically significant difference in clinical pregnancy rate among groups(P=0.133).Conclusion Performing IUI in PMSC (0.15-60.00)×106 after processing can get preferable pregnancy rates.

Objective To investigate the clinical efficacy of Dahuang Xiaozhi Suppository combined with tetracycline in treating type Ⅲ prostatitis infected with nano-bacteria. Methods Totally 120 patients of type Ⅲprostatitis infected with nano-bacteria were randomly divided into treatment group and control group, with 60 cases in each group. Both groups were disabled anti-infective drugs and other preparations for diet and life intervention. Both groups received tetracycline, once a tablet, twice a day, orally. The treatment group received Dahuang Xiaozhi Suppository, once a capsule, once a day, placing in the anus 3-4 cm. 10 d was a treatment course, for 3 courses. The clinical efficacy, major symptoms improving time, NIH-CPSI, leukocyte count and ecithin corpuscles in expressed prostatic secretion (EPS), urinary flow rate, and cytokine content of TNF-α, IL-6, IL-8, were evaluated. Results The total effective rate was 100.0% (60/60) in treatment group and 83.3% (50/60) in the control group, and treatment group was higher than the control group (P<0.05). Pelvic pain, urinary symptom, and scrotum wet improvement time of treatment group were lower than the control group (P<0.05). Compared with before the treatment, the NIH-CPSI pain scores, urinary symptom scores, life quality score, leukocyte count and TNF-α, IL-6, IL-8 content of EPS in both groups were significantly lower after treatment (P<0.05), and the cases with lecithin corpuscles +++ - ++++ in both groups significantly increased after treatment (P<0.05). The urinary peak flow rate and mean flow rate in both groups were higher after treatment (P<0.05). There was statistical significance in the scores of NIH-CPSI pain, urinary symptom and life quality, leukocyte count and TNF-α, IL-6, IL-8 content of EPS, and the cases with lecithin corpuscles ++++ between the two groups (P<0.05). Conclusion Dahuang Xiaozhi Suppository can improve the efficacy of treating type Ⅲ prostatitis infected with nano-bacteria. Combining Dahuang Xiaozhi Suppository with tetracycline can reduce prostate inflammation, with obvious efficacy.