Objective To analyze the effect of inducible costmulator (ICOS)/inducible costmulator ligand (ICOSL) signaling pathway on hepatic fibrosis in mice infected with Schistosoma japonicum.Methods Seventy-eight ICOSL knockout (ICOSL-KO) mice and 77 wild type C57BL/6J mice were used as experimental schistosomiasis model infected with Schistosoma japonicum.The sera of mice were collected on the day before infection (0 week),and at 4,7,12,16 and 20 weeks post infection.Then,the concentrations of hyaluronic acid (HA) and hydroxyproline (HYP) in mice sera were measured by sandwich enzyme linked immunosorbent assay (ELISA) kits.The expressions of transforming growth factor β1 (TGF-β1),α-smooth muscle actin (a-SMA) and Collagen-Ⅰ in livers from ICOSL-KO/wild type mice were assessed by immunohistochemical staining.The granulomatous pathology and fibrosis level in mice liver were dynamically observed by hematoxylin and eosin (HE) staining and Masson's staining,respectively.The difference between groups was detected by t test or x2 test when appropriate.Results After infection with Schistosoma japonicum,the levels of HA and HYP were gradually increased.In ICOSL-KO mice,the levels of HA at 7,12,16 and 20 weeks post infection were all significantly lower than those in wild type mice [(161.32±15.44) vs (186.01±21.24) ng/mL,t=2.528 2,P＜0.05; (166.73±18.18) vs (231.39±20.12) ng/mL,t=4.342 4,P＜0.05; (193.58±21.06) vs (252.51±25.29) ng/mL,t=4.003 9,P＜0.05; (253.98±24.53) vs (310.88±23.86) ng/mL,t=3.718 0,P＜0.05].Similarly,HYP levels in ICOSL-KO mice at 12,16 and 20 weeks post infection were all significantly lower than those in wild type mice (all P＜0.05).Immunohistochemical staining showed that TGF-β1,α-SMA and Collagen-Ⅰ expressions in liver of ICOSL-KO mice from 7 to 20 weeks post infection were all significantly lower than those of wild type mice (all P＜0.05).HE staining showed,the volume of liver egg granulomas of ICOSL-KO mice was significantly smaller than that of wild type C57BL/6J mice (P＜0.01).Furthermore,Masson's staining showed that the level of hepatic fibrosis in ICOSL-KO mice was lower than that in wild type mice and the fibrosis scores were statistically different between two groups (all P＜0.05).The mortality rate of the wilde type C57BL/6J mice was higher than that of ICOSL-KO mice.After 20 weeks of infection,the difference was statistically significant (55.84 ％ vs 37.18 ％,x2 =5.427,P＜0.05).Conclusions The degree of hepatic fibrosis and related indicators are obviously down-regulated in ICOSL-KO mice infected with Schistosoma japonicum.These findings suggest that ICOS/ICOSL signaling pathway has an important impact on the process of hepatic fibrosis caused by Schistosoma japonicum.

Objective:To analyze effect on the CD154-CD40 signaling pathway and Th1/Th2 polariza-tion by deficient inducible co-stimulator ( ICOS)-ICOS ligand ( ICOSL) signaling in mice infected with Schistosoma japonicum. Methods:ICOSL knockout ( ICOSL-KO) mice and wild-type C57BL/6J mice were used as experimental Schistosomiasis model infected with Schistosoma japonicum. The expressions of CD154 and CD40 on splenocytes and on inflammatory cells around granulomatous infiltration of liver in mice infected with Schistosoma japonicum were analyzed by flow cytometry,immunohistochemical staining, respectively, on the day before infection (0 week) and at the end of 4, 7, 12, 16 and 20 weeks post-infection. The splenocytes of the mice were stimulated with soluble egg antigen( SEA) for 72 hours, then the concentrations of interferon gamma(IFN-γ) and interleukin-4 (IL-4) in the culture supernatants were measured by sandwich enzyme-linked immunosorbent assay ( ELISA) kits. The levels of SEA-specific an-tibodies of IgG and IgG1 and IgG2a were measured in the mice sera by ELISA. The granulomatous pa-thology in the mice liver was dynamically observed by hematoxylin and eosin ( HE ) staining. Results:Compared with the wild-type C57BL/6J mice, the expressions of CD154 on CD4 + T splenocytes [(18. 62 ± 4. 76)% vs. (27. 91 ± 3. 94)%, (22. 44 ± 4. 67)% vs. (40. 86 ± 5. 21)%, (25. 50 ± 6. 81)% vs. (43. 81 ± 8. 41)%, (20. 22 ± 5. 28)% vs. (40. 95 ± 7. 34)%, (17. 87 ± 4. 59)% vs. (33. 16 ± 6. 31)%, all P <0. 01] and of CD40 on CD19 + B splenocytes [(19. 43 ± 3. 26)% vs. (24.37 ±3.59)%, (23. 00 ± 4. 47)% vs. (31. 80 ± 5. 86)%, (24. 46 ± 5. 01)% vs. (35. 85 ± 5. 32)%, (23. 42 ± 4. 69)% vs. (33. 30 ± 6. 14)%, (22. 85 ± 3. 78)% vs. (30. 88 ± 5. 94)%, all P<0 . 05 ] in the ICOSL-KO mice significantly decreased at the end of 4 , 7 , 12 , 16 and 20 weeks post-infection. Moreover, the expressions of CD154[(0. 319 ± 0. 066) vs. (0. 488 ± 0. 086), (0. 389 ± 0. 067) vs.(0.596±0.082),(0.378±0.064) vs.(0.543±0.072),(0.348±0.069) vs.(0.523±0.076), all P<0. 01] and CD40[ (0. 398 ± 0. 066) vs. (0. 546 ± 0. 079), (0. 461 ± 0. 085) vs. (0. 618 ± 0. 076), (0. 453 ± 0. 087) vs. (0. 587 ± 0. 074), (0. 449 ± 0. 065) vs. (0. 565 ± 0. 082), all P <0 . 05 ] on inflammatory cells around granulomatous infiltration in liver from the ICOSL-KO mice were sig-nificantly lower than those of the wild-type C57 BL/6 J mice at the end of 7 , 12 , 16 and 20 weeks post-in-fection. The levels of IFN-γ of the ICOSL-KO mice were significantly higher than those of the wild-type C57BL/6J mice at the end of 4, 7, 12, 16 and 20 weeks post-infection (P <0. 05). However, the levels of IL-4 of the ICOSL-KO mice were significantly lower than those of the wild-type mice ( P <0. 05). Compared with the wild-type C57BL/6J mice, the levels of SEA-specific antibodies of IgG and IgG1 and IgG2a in the sera of the ICOSL-KO mice significantly decreased (P<0. 01). Moreover, The Th2 differentiation index of the ICOSL-KO mice was significantly lower than that of the wild-type mice in post-infection (P<0. 01). Also, the ratio of IgG1/IgG2a of the ICOSL-KO mice were significantly lower than that of the wild-type mice at the end of 7 , 12 and 16 weeks post-infection ( P<0 . 05 ) . And the vo-lume of liver egg granulomas of the ICOSL-KO mice was significantly smaller than that of the wild-type mice ( P <0 . 01 ) . Conclusion: These findings suggest that there is obvious down-regulation in the expressions of CD154 and CD40 and impairment of Th2 immune response in the ICOSL-KO mice infected with Schistosoma japonicum, accompanying with notedly reduced hepatic granulomatous pathology. The ICOS-ICOSL signaling has a regulatory effect on CD154-CD40 signaling pathway, and may play an impor-tant role in the hepatic egg granuloma formation of Schistosomiasis.

Objective To investigate the protective effect of resveratrol ( RES) against hydrogen peroxide ( H2 O2 )-induced oxidative injury to astrocytes and the related mechanism. Methods Subcultured astrocytes were randomly divided into four groups:negative control group ( treated with normal culture medium) , model control group ( treated with 100 μmol·L-1 H2 O2 for 12 h), resveratrol low dose group (treated with 20 μmol·L-1 RES for 24 h H2O2 for 12 h) and resveratrol high dose group ( treated with 40 μmol·L-1 RES for 24 h before H2 O2 for 12 h) . Cell viability was detected by MTT assay, apoptosis rate was detected by flow cytometry, apoptotic cell morphology was detected by hochest33258 staining, and the expression of apoptosis-related factors such as caspses-3 and caspase-9 were measured by colorimetric detection. Results MTT assay showed that after treatment with 5, 10, 20, and 40 μmol·L-1 RES for 24 h, cell viability was (100. 46±3. 17)%, (101. 33± 3.14)%, (101. 33±1. 30)%, and (99. 67±2. 62)%, respectively, and the difference was not statistically significant as compared with the negative control group [(98. 33±2. 13)%, P>0. 05]. RES showed no effect on astrocyte activity, after treatment with 20 and 40 μmol·L-1 RES, astrocyte activity was significantly elevated to (54. 67±4. 11)% and (70.33± 2. 61)% as compared with model control group (t=3. 59, 7. 13, P<0. 05), RES inhibited hydrogen peroxide-induced decrease in cell viability. Flow cytometry results showed that after treatment with 20, 40 μmol·L-1 RES, the apoptosis rate of astrocytes significantly decreased to (35.51±3. 56)% and (14. 12%±3. 19)% (t=4. 26, 6. 33, P<0. 01) as compared with model control group (46. 31±4. 16)%. Hochest 33258 staining showed that RES inhibited hydrogen peroxide-induced cell apoptosis, besides, the RES treatment also could reduce H2 O2-induced expression of caspses-3 and caspase-9 in astrocytesin a time-dependent manner. Conclusion RES can inhibit hydrogen peroxide-induced astrocytes apoptosis through inhibiting the expression of caspses-3 and caspase-9, which can provide experimental evidence for its treatment of central nervous disorders.

Objective To investigate the efficacy and safety,as well as surgical essentials of upper urinary tract calculi (calculi being≤2 cm in diameter) complicated with pre-surgical uncertain pyonephrosis through one-stage renal calculi removal by percutaneous nephrolithotomy and suction device.Methods Retrospective analysis was used to detect the thirty-three cases with upper urinary tract calculi complicated with pre-surgical uncertain pyonephrosis collected from August 2010 to March 2016 in Yellow River Sanmenxia Hospital Affiliated to Henan University of Science and Technology,all the cases in the group had no pre-surgical fever,no apparent infection by blood-urine routine test,different degrees of hydronephrosis and no indications of pyonephrosis confirmed by CT and color Doppler ultrasonography,no pre-surgical anti-infection cure,pyonephrosis was found during the operation.First of all,a suction device was used to suck pus through percutaneous renal channel,rinsing repeatedly with small amount of fluid until the sucked rinsing fluid was clear;then,the one-stage calculi was removed by percutaneous renal lithotripsy,and the pus was sent to be cultured during the surgery,and the cases were treated by postoperative intravenous anti-infection for seven to seventeen days.Operation condition,postoperative blood routine,temperature,calculi removal and other clinical recovery conditions should be carefully observed.Results All the 33 cases underwent the one-stage single channel percutaneous nephrolithotom,the surgery was successful,the operation lasted 28-59 minutes,with an average of 41 minutes per case.Within the first-week of operation,CT reexamination showed the stone-free rate was 90.9% (30/33),and three cases had residual stone,the maximum diameter was about 6mm;seven cases had high fever within 3 days after the surgery (21.2%),and the temperature in four cases was higher than 38.5℃(12.1%) and two cases had fever within 3-6 days after surgery (6.1%),the highest temperature reached 38.0℃,and no fever existed after 6 days.Compared with preoperative values,the postoperative blood routine indicated that the surgery-relevant hemoglobin has decreased to (6.16±5.21) g/L;three days after surgery,white blood cell count was (7.16±4.86) 109/L.There were no severe complications such as sepsis,septic shock,renal abscess,hemorrhage.All cases were followed up for 6 to 36 months and no secondary pyonephrosis or renal dysfunction occurred during that period.Conclusion The treatment of upper urinary tract calculi,complicated with pyonephrosis without pre-surgical fever through one-stage renal calculi removal by percutaneous nephrolithotomy and suction device is effective and safe,it can be used as the routine method in basic hospitals and more attention should be paid to the operation and renal pelvic pressure in order to avoid the occurrence of complication.

Objective To investigate the expression and clinical significance of UL16 binding protein 3 (ULBP3) in human esophageal squamous cell carcinoma (ESCC) and its correlation with nature killer (NK) cells.Methods The relative expression of ULBP3 in the ESCC tissues and corresponding carcinoma adjacent tissues of 40 patients was detected by realtime-poly merase chain reaction (PCR),immunohistochemical staining and Western blot methods.The percentage of NK cells in peripheral blood of same patients was examined by flow cytometry.The correlation between ULBP3 and the percentage of NK cells was analyzed with Pearson method.Results The expression of ULBP3 at mRNA level in the tumor tissues ([4.96 ±-6.11]×10-3) was significantly higher than that of corresponding carcinoma adjacent tissues ([1.64 ± 2.96]× 10-3,t =3.656,P＜ 0.01).The immunohistochemical staining results indicated that the positive rate of ULBP3 in the tumor tissues was 60％ (24/40),however that of corresponding carcinoma adjacent tissues was only 32.5％(13/40,t=3.921,P＜0.01).The Western blot results indicated that the expression of ULBP3 at protein level in the tumor tissues was significantly higher than that in the corresponding carcinoma adjacent tissues.The relative expresssion ULBP3 at mRNA level in carcinoma tissues of ESCC patients with lymph node metastasis and at TNM stage Ⅲ was higher than that of ESCC patients without metastasis and at TNM stage Ⅰand Ⅱ (t=4.839,4.192,P＜0.05).There was no significant correlation between the expression and ages,gender,location of tumors and the differentiation degree of tumor (P＞0.05).At early and mid stage of the tumor,the expression of ULBP3 at mRNA level was positively correlated with the percentage of NK cells in peripheral blood (r=0.5233,P＜0.05),however there was no correlation at advanced stage.Conclusion ULBP3 was highly expressed in ESCC and may be involved in the immune regulation of NK cells.

Objective To measure the activity energy expenditure(AEE) of healthy adults during level walking by using indirect calorimetry,and to analyze the characteristics and underlying influencing factors such as age and gender. Methods A total of 60 healthy adults aged 20-50 years (30 males and 30 femdes) participated in the study.All the subjects were divided into six groups by gender and age (the age span of each group was 10 years). The subjects were arranged to walk at speeds of 3.5,4.5,5.5km/h and run at 5.5,6.5,7.5km/h, respectively, on the treadmill. The resting energy expenditure ( REE ) and AEE were measured during walking and running at different speeds.There was a 5-minute rest among the test sessions. Results No difference in terms of AEE between the female and male at the same age ( P ＞ 0. 05 ). During 3.5km/h walking,AEE of 21-30 year-old females was lower than 31-40 yearold females and males( P ＜0.05 ) ;During 4.5km/h walking,AEE of 31-40 year-old females was higher than 21-30 yearold females and males and 41-50 year-old females; AEE of 21-30 year-old females was lower than 41-50 year-old males (P ＜ 0.05 ) ;During 5.5km/h walking,AEE of 31-40 year-old females was higher than 21-30 year-old females and males ( P ＜ 0. 05 ). During 6.5 km/h running, AEE of 31-40 year-old females was higher than 41-50 year-old females ( P ＜0.05), while during 7.5km/h running, AEE of 21-30 year-old males was higher than 41-50 year-old females (P ＜0.05 ). It was also found that the AEE of all groups except the 41-50 year-old females group was higher when walking at the speed of 5.5km/h than running at the same speed( P ＜ 0.05 ). Conclusions Age has more effect on REE and AEE than the gender. AEE of elder subjects is higher than that of the younger ones during walking, however, AEE of younger people increases faster than the elders during running. AEE of 31-40 year-old females is the highest in all groups both in walking and running. AEE in running is higher than in walking at the same speed.

Objective To study the expression levels of inducible costimulator (ICOS) on CD4+ T cells in peripheral blood in patients with chronic hepatitis B and its change after treated with interferon.Methods All 56 patients were divided into two groups (interferon group has 28 cases,lamivudine group has 28 cases),and interferon group treated by PEG-IFN-α 2a,lamivudine group treated by lamivudine,respectively.There were 20 healthy individuals as control group.The levels of ICOS on CD4+ T cells of patients with chronic hepatitis B were kinetically detected by flow cytometry.The copies of HBV DNA in sera were dynamically detected by real-time PCR.Results The levels of ICOS on CD4+ T cells in patients with chronic hepatitis B was higher than that of normal controls(P＜0.001 ).However,the expressions of ICOS on CD4+ T cells in patients could be deduced by PEG-IFN-α 2a and obviously decreased after treatment.There was significant difference between interferon group and lamivudine group (P＜0.05 in 24 weeks,and P＜0.01 in 48 weeks).After treatment,the change values of ICOS in interferon group was positively correlated with the change copies of HBV DNA (r =0.972,P＜0.001 ).However,the change values mentioned above that did not find correlation in lamivudine group(r=-0.101,P=0.608).Conclusion The study shows the patients with chronic hepatitis B have disordered in cellular immunity and increased with the expression of ICOS on CD4+ T cells.PEG-IFN-α 2a could decrease the expression of ICOS on CD4+ T cells in patients,and correct the Th2 type immune polarization to some extent,and that plays a positive role in antiHBV.

Objective To investigate the effects of metformin on plasma ghrelin and peptide YY (PYY)levels in newly-diagnosed type 2 diabetic patients,and to study the impact of metformin on body weight.Methods A prospective nested case-control study was designed as a research protocol.Sixty four newly-diagnosed type 2 diabetic patients were treated with metformin for 12 weeks.The patients were divided into two groups:weight loss group and non-weight loss group according to the changes in body weight after metformin treatment.Fasting plasma ghrelin and PYY levels and other metabolic parameters were measured before and after metformin treatment.ResultsFasting plasma ghrelin level was significantly decreased in the patients after metformin treatment [ ( 10.71 ±2.68 vs 11.81 ±3.19 )ng/ml,P＜0.05 ].Fasting plasma PYY level was significantly increased in patients after metformin treatment [ ( 136.86+39.14 vs 128.42+37.31 ) pg/ml,P＜0.05 ].After metformin treatment,43.7％ of the patients lost body weight significantly.Fasting plasma ghrelin level was decreased by 16.6％ after treatment in the weight loss group,as compared with 6.2％ in non weight loss group( P＜0.05 ).Fasting plasma PYY level was increased by 10.8％ after treatment in the weight loss group,as compared with 3.5％ in the non-weight loss group (P ＜ 0.05 ).Conclusions The fasting plasma ghrelin level in the weight loss group was lowered more significantly compared with that in the non-weight loss group after metformin treatment.The fasting plasma PYY level in the weight loss group was elevated more significantly as compared with that in the non-weight loss group after metformin treatment.The mechanism remains to be further studied.

AIM: To observe the therapeutic effect of hepatocyte growth promoting factor (PHGF) combined with transmetil in treatment of chronic severe hepatitis (CSF). METHODS: 120 patients were divided randomly into two groups: the combined treatment group and the routine group. All the patients received ordinary therapy, and 62 patients, based on the therapy, received PHGF combined with transmetil in the combined treatment group. RESULTS: Compared with the routine treatment group, the levels of total serum bilirubin and prothrombin time were decreased significantly (P

Background and purpose:BCRP was recently discoveried as a membrane transport protein affiliated with multiple drug resistance(MDR).The aim of present study was to investigate the expression of breast cancer resistance protein(BCRP) in primary breast carcinoma and its potential significance of guiding breast carcinoma chemotherapy and to determine whether such expression can be used as a predict factor for chemosensitivity.Methods:Expression of BCRP in 31 primary breast carcinoma tissues was determined by flow cytometry.The relationship between the expression of BCRP with the clinicpathological characteristics and the prognosis of breast carcinoma patients was also analyzed.Results:The expression level of BCRP was higher in breast carcinoma tissue(0.282581?0.183686) than control group(0.03125?1.000905).There was no statistical difference befween BCRP and state of ER,PR,C-erbB_2 and EGFR in breast carcinoma tissues.In addition,it was the same situation no matter whether axillary lymph node was metastatasied or not.Conclusions:The results suggest that BCRP is expressed in primary breast carcinoma and is a cell membrane protein independent on ER,PR,c-erb-B-2,EGFR.Based chemotherapy is more effective for the patients with the overexpression of BCRP in primary breast carcinoma tissue.