Objective To investigate the therapeutic effects of curcumin(Cur)on trinitrobenzene sulphonic acid(TNBS)induced colitis and to investigate cytokines change in colon mucosa,spleen and sera.Methods Colitis was induced in SD rats by intrarectal injection of TNBS(100 mg/kg).Experi- mental animals were divided into negative control group,TNBS group and Cur therapeutic group(Cur, 30 mg?kg~(-1)?d~(-1),intraperitoneal injection).Expression of cytokines mRNA in colon mucosa was observed by RT-PCR,intracellular cytokines of interferon(IFN)-?and interleukin(IL)-4 in splenocytes were detected by flow cytometry(FCM),concentrations of IFN-?and IL-4 in sera were determined by enzyme-link immunosorbent analysis(ELISA).Results After treatment with Cur,macroscopic scores (3.9?1.0 vs.2.2?0.7),myeloperoxidase(MPO)activity(15.0?2.6 vs.7.3?1.4),mRNA of IFN-?(1.02?0.07 vs.0.06?0.02,mRNAof IL-12(0.29?0.05 vs.0.11?0.01)and the ratio of IFN-?/IL-4(11.44?0.97 vs.0.38?0.10)in colon mucosa,proportion of IFN-?CD4~+(31.7?7.5 vs. 21.1?3.7)and the ratio of IFN-?/IL-4 CD4~+(19.9?5.1 vs.6.1?1.8)in splenocytes,concentra tions of IFN-?[(1528?159)pg/ml vs.(513?14)pg/ml] and the ratio of IFN-?/IL-4(19.5?4.1 vs. 4.2?0.6)in sera were all decreased(P＜0.05 or P＜0.01).Meanwhile,mRNA of IL-4(0.09?0.01 vs.0.15?0.04)and IL-10(0.28?0.08 vs.0.63?0.12)in colon mucosa,proportion of IL-4 CD4~+ (1.6?0.5 vs.3.4?1.1)in splenocytes and concentrations of IL-4 in sera[(81?15)pg/ml vs.(124?20) pg/ml] were all increased after the treatment of Cur(P＜0.05 or P＜0.01).Conclusions Cur showed ther- apeutic effects on TNBS-induced colitis,and the mechanism might be through regulating the balance of Th1/ Th2 in colon locally and systematically.

OBJECTIVE:To develop a method for simultaneous determination of chlorogenic acid,geniposide,gentiopicro-side,ferulic acid,baicalin and ammonium glycyrrhizinate in Yindan pinggan capsule. METHODS:HPLC method was adopted. The separation was performed on Wonda SilTM-C18 column with mobile phase consisting of acetonitrile-0.4% phosphoric acid solu-tion(gradient elution)at the flow rate of 0.8 mL/min. The detection wavelength were set at 325 nm(chlorogenic acid,ferulic ac-id),250 nm (geniposide,ammonium glycyrrhizinate) and 275 nm (gentiopicroside,baicalin). The column temperature was 30 ℃. RESULTS:The linear ranges were 0.087-3.480 μg for chlorogenic acid(r=0.9998),0.201-8.040 μg for geniposide(r=0.9997),0.200-8.000 μg for gentiopicroside(r=0.9995),0.016-0.640 μg for ferulic acid(r=0.9999),0.105-4.200 μg for ba-icalin (r=0.9999) and 0.028-1.120 μg for ammonium glycyrrhizinate (r=0.9995),respectively. The limits of quantitation were 1.31,0.75,1.14,1.25,0.94,0.98 ng,and the limits of detection were 0.87,0.67,0.96,0.93,0.60,0.88 ng,respectively. RSDs of precision,stability and reproducibility tests were lower than 2.0%;recoveries were 99.9%-101.9%(RSD=0.7%,n=6), 98.7%-100.9%(RSD=0.9%,n=6),98.1%-101.5%(RSD=1.4%,n=6),98.5%-101.3%(RSD=1.3%,n=6),98.5%-101.7%(RSD=1.2%,n=6),98.2%-101.4%(RSD=1.2%,n=6),respectively. CONCLUSIONS:The method is simple and accurate , can be used for simultaneous determination of chlorogenic acid,geniposide,gentiopicroside,ferulic acid,baicalin and ammonium glycyrrhizinate in Yindan pinggan capsule.

Objective To study the risk factors of mortality in severe sepsis and septic shock patients. Methods 142 patients with severe sepsis and septic shock in ICU were observed and divided into alive group (98 patients)and dead group (44 patients)by using hospital mortality.The risk factors of mortality in severe sepsis and septic shock patients were assessed by binary logistic regression.Results Independent mortality risk factors were inotropic agents (OR =4.329,95%CI:1.045 -17.937,P =0.043),blood glucose >10 mmol/L (OR =3.771,95%CI:1.214 -11.710,P =0.022)and APACHE Ⅱ score (OR =3.098,95%CI:2.012 -4.760,P =0.000),while PaO2 /FiO2 after early goal -directed therapy (EGDT)was protective factor (OR =0.682,95%CI:0.500 -0.930, P =0.016).Conclusion Severe sepsis and septic shock patients with inotropic agents,blood glucose >10mmol/L, high APACHE Ⅱ score and decreased PaO2 /FiO2 after EGDT indicate poorly prognosis.

Arachnoid ; pathology ; surgery ; Bone Neoplasms ; surgery ; Humans ; Male ; Middle Aged ; Neurilemmoma ; surgery

Objective To determine whether Fudenine is a novel membrane protein. Methods Green fluorescence protein(GFP)was used to localize Fudenine in vivo. GFP, as a control, was targeted to cytoplasm.Epithelial cell, CBRH7919, and non-epithelial cell, L-6TG, were cultured and transiently transfected by using the lipofectamine reagent. After 48 h, intact cells were examined with fluorescence microscope for Fudenine. Results Reporter plasmid pEGFP-N1, as a control , was expressed and localized to cytoplasm. But Fudenine, driven by the cytomegalovirus promoterenhancer contained in the pEGFP-N1 vector, was overexpressed and targeted to cellular membrane. Conclusions Fudenine is a novel membrane protein. It may play the similar role with its homologues AC133 antigen and prominin in human and mouse.respectively. It might be involved in signaling transduction and regulate blood glucose metabolism in vivo.

OBJECTIVETo study the therapeutic mechanism of Zhizhu Pill (ZP) for treating functional dyspepsia (FD) rats.

METHODSTotally 30 ten-day-old male rats were randomly divided into the normal control group (n =10) and the model group (n = 20). The FD rat model was induced using gastric administration of 0.1% iodoacetamide (IA) combined tail clamping. The model was evaluated when rats were 8-week old. Successfully modeled rats were randomly divided into the model group (n = 10) and the ZP group (n = 10). Rats in the normal group and the model group were administered with normal saline by gastrogavage, while those in the ZP group were administered with ZP Decoction (2 mL/100 g) by gastrogavage. All medication lasted for 7 successive days. The contractile activity in in vitro longitudinal gastric muscle was recorded using Power Lab biological signal collecting system. The expression of growth hormone secretagogue receptor (GHSR) in stomach of FD rats was detected using Western blot and immunohistochemistry (IHC).

RESULTSCompared with the normal group, average frequencies of gastric contraction and changing rates of amplitude obviously decreased in the model group (P < 0.05). Results of Western blot and IHC showed that the expression of GHSR decreased in the model group (P < 0.01). Compared with the model group, average frequencies of gastric contraction and changing rates of amplitude obviously increased in the ZP group (P < 0.05). Results of Western blot and IHC showed that the expression of GHSR increased in the ZP group (P < 0.01).

CONCLUSIONZP could promote the gastric motility in FD rats induced by gastric administration of IA combined tail clamping, and its mechanism might be related to up-regulating GHSR protein level.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Dyspepsia ; drug therapy ; Gastrointestinal Motility ; Male ; Muscle Contraction ; drug effects ; Muscle, Smooth ; drug effects ; metabolism ; Random Allocation ; Rats ; Receptors, Ghrelin ; metabolism

OBJECTIVETo investigate the clinical effects of plate and xenogenic bony plate for the treatment of nonunion of femoral shaft fracture after initial fixation with locked intramedullary nailing.

METHODSFrom February 2006 to June 2013, 21 cases with nonunion of femoral shaft fracture after initial fixation with locked intramedullary nailing were treated with femoral plate and contralateral xenogenic bony plate. There were 12 males and 9 females with an average age of 34.8 years old (ranging from 18 to 62 years). The time of nonunion was 9 to 18 months (averaged 12.8 months). The clinical outcomes of the treatment were evaluated by Merchan assessment.

RESULTSAll of the patients were primary healing, and on complications such as infection,fat embolism, internal fixation breaking or rotational deformity, shortening were occurred. All the cases were followed up for 13.2 months (ranging from 8 to 24 months). Nineteen cases were bone healed,the time of union averaged 6.2 months (ranging from 4 to 9 months). Two cases appeared delayed union and gained bony union after 7 to 13 months' observation. According to the criterion of Merchan,the results were excellent in 13 cases, good in 6 cases, poor in 2 cases.

CONCLUSIONTreatment of nonunion of femoral shaft fracture after initial locked intramedullary fixation with plate and xenogenic bony plate has advantages of high curative rate and low complications, good postoperative function recovery, it is a reliable treatment to elevate the stability of fixation and promote the osteogenesis.

Adolescent ; Adult ; Bone Plates ; Female ; Femoral Fractures ; surgery ; Fracture Fixation, Intramedullary ; methods ; Fractures, Ununited ; surgery ; Humans ; Male ; Middle Aged

Objective To observe the influence of different doses of intravenous immunoglobulin (IVIG) on function of T cells in cord blood of premature infants and to explore the immunomodulating mechanism of IVIG.Methods Cord blood mononuclear cells (CBMC) were isolated by density gradient centrifugation method from 15 preterm infants born between June 1,2011 to December 31,2011 at 32 34 gestational weeks.Three groups were formed according to the concentrations of IVIG used for CMBC culturing in vitro (Group A,6 mg/ml; Group B,9 mg/ml; Group C,12 mg/ml).After 72 hours in sterile conditions,the levels of IFN 7,IL 4,IL-10 and TGF β1 in the supernatant were determined by enzyme-linked immunosorbent assay.The expression of CD4+ CD25+ Foxp3+ Treg cells was examined by fluorescent activated cell sorter.Differences among groups were compared by one way analysis of variance.For comparison between two groups,LSD test was used.Results (1) The level of IFN γ secreted by CBMC in Group C was 0.03 ± 0.02,which was much lower than that in Group A(0.18±0.08) and Group B(0.13±0.05) (F=5.72,both P＜0.01).The level of IFN-γ in Group B and Group A did not show statistical difference (P＞0.05).Compared with Group A(0.03±0.01),the level of IL4 was much lower in Group B (0.02±0.01) and Group C(0.01±0.01) (F=20.38,both P＜0.01),while no significant difference was shown between Group B and Group C (P＞0.05).(2) No significant difference was found in the expression of CD4+CD25+Foxp3+Treg cells among different groups (F 0.67,P＞0.05).(3) The level of IL 10 in Group C was 3.02 ± 3.79,which was significantly lower than that in Group A (10.78±5.44) and Group B (6.90±4.64)(F=4.68,P＜0.01 and 0.05).The level of IL-10 inGroup B was still lower than that in Group A (P＜0.05).The levels of TGF-β1 in Group C(8.44± 13.71) and Group B(16.15 ±13.94) were significantly lower than that in Group A(30.23 ± 16.32) (F=5.22,P＜0.01,P＜0.05),but there was no significant difference between Group B and Group C (P＞0.05).Conclusions IVIG might inhibit the function of Th cells in CBMC of preterm infants in a dose dependent manner.IVIG could inhibit the function of natural Treg cells by regulating the secretion of cytokines IL-10 and TGF-β1 in a dose dependent effect.

Objective To investigate the effects of donor-specific regulatory T cells (Treg) transfusion on islet allograft survival. Methods Allogeneic fresh islets from Balb/c mice were transplanted to streptozotocin-induced diabetic C57 mice. The survival of islet allografts was observed. The experiment was divided into 3 groups: control group, nothing had been done to the recipients; simple islet transplantation group, the recipients received the islet transplantation only; experimental group, the recipients were given 1 ×106 Treg, then received islet transplantation. Results Blood glucose (BG) was above 16. 7 mmol/L after islet transplantation in control group; In simple islet transplantation group,BG level returned to normal level 1 to 2 days after transplantation, and hyperglycemia appeared 7 to 11 days after transplantation and maintained as the same as that before transplantation; In experimental group, BG level returned to normal level 2 days after transplantation and maintained at a low level,and at the 21st day after transplantation BG level was over 16. 7mmol/L in some recipients. Islet allograft survival in experimental group was significantly prolonged as compared with simple islet transplantation group. Conclusion Donor-specific Treg transfusion could prolong the islet allograft survival,and maybe have positive effect on tolerance induction of islet transplantation.

Objective To assess whether asymmetric spin echo version of echo planar imaging (ASE EPI) sequence can reflect cerebral oxygen metabolism by means of the changes in oxygen extraction fraction values before and after surgical operations in the canine model of ischimia. Methods A total of 12 healthy crossbreed dogs were enrolled in this study. All canines underwent cerebral MR imaging including coronal T2WI, DWI, MRA, ASE EPI and 3D PCASL. The bilateral carotid arteries (CA) were separated by surgical operations. The bilateral CA were ligated in 6 dogs, and the other 6 dogs had embolization in ligated bilateral CA. The cerebral MR imaging with the above protocol was repeated at 1 hour after surgical operation for each dog. All dogs were executed when they finished the MR examinations. Then the whole brains of the dogs were taken out for cutting coronal sections and 2, 3, 5-triphenyltetrazolium chloride (TTC) staining. The ASE EPI data were processed by the Functool software in the workstation to generate OEF maps. For measuring the OEF values before and 1 hour after surgical operations, one and the same region of interest (ROI) was respectively placed in the left and right hemispheres on the OEF maps for each dog. 3D PCASL data were processed by the Functool software in the workstation to yield CBF maps. The method for measuring the CBF values before and 1 hour after surgical operations on CBF maps was same as that for measuring on OEF maps. The changes of measurements before and after surgical operations were compared between the dogs with ligated bilateral CA and the dogs with embolization in ligated bilateral CA by using an independent samples t test. The measurements before and after surgical operations were respectively compared in the dogs with ligated bilateral CA and the dogs with embolization in ligated bilateral CA by using a paired t-test. The cutting slices with TTC staining were analysed for determining ischimia. Results The CBF values of the dogs with ligated bilateral CA were (59.81±23.59) and (38.56± 12.98) ml/(min · 100 g) before and after surgical operations, respectively. The CBF values of the dogs with embolization in ligated bilateral CA were (58.94±18.35) and (28.01±11.41) ml/(min·100 g) before and after surgical operations, respectively. The CBF values were significantly lower before surgical operations than after surgical operations in the dogs with ligated bilateral CA and the dogs with embolization in ligated bilateral CA, respectively (t=8.92 and 13.42, respectively; P=0.00, respectively). The CBF values were decreased more significantly in the dogs with embolization in ligated bilateral CA than in the dogs with ligated bilateral CA (t=2.92, P=0.00). The OEF values of the dogs with ligated bilateral CA were 0.29±0.02 and 0.32 ± 0.06 before and after surgical operations, respectively. The OEF values of the dogs with embolization in ligated bilateral CA were 0.29 ± 0.02 and 0.35 ± 0.06 before and after surgical operations, respectively. The OEF values were significantly higher before surgical operations than after surgical operations in the dogs with ligated bilateral CA and the dogs with embolization in ligated bilateral CA, respectively (t=-7.21 and-4.43, respectively;P=0.00, respectively). The OEF values were increased more significantly in the dogs with embolization in ligated bilateral CA than in the dogs with ligated bilateral CA (t=2.18, P=0.03). The pathological results showed that the dogs with embolization in ligated bilateral CA had swollen cortex with focal infarction which was matched with the hyperintensity areas on the OEF and DWI images. However, the dogs with ligated bilateral CA only had swollen cortex whereas didn't have focal infarction. Conclusions ASE EPI sequence can reliably reflect cerebral oxygen metabolism in ischemia. This sequence has an important role for assessing hemodynamic state in patients with cerebral vascular diseases.