Objective:To study the effects and mechanisms of acetyl-11-keto-β-boswellic acid ( AKBA) in myocardial ischemic model induced by isoproterenol hydrochloride ( ISO) in rats. Methods:The SD rats were randomly divided into the sham group, model group, AKBA low dose group and AKBA high dose group. Myocardial injury model was induced by subcutaneous injection of ISO (100 mg·kg-1 ) . The change of ST segment in ECG was observed. Creatine kinase ( CK-MB) , cardiac troponin I ( cTnI) , lactate dehydro-genase( LDH) , malondialdehyde( MDA) and superoxide dismutase( SOD) in the blood were detected by ELISA. The change of histo-logical tissue was determined by HE staining, and cell apoptosis was analyzed by TUNEL assay. Results: Serum CK-MB, cTnI and LDH were decreased significantly in AKBA high dose group when compared with those in the model group. Compared with that in the model group, MDA content was lowered and the SOD activity was increased in AKBA high dose group. Furthermore, AKBA high dose group improved the pathologic changes of myocardium. TUNEL assay revealed significant reduction of cardiomyocytes apoptosis in the hearts of the ischemic rats in AKBA high dose group. Conclusion:AKBA has excellent cardioprotective effect on myocardial ischemic induced by ISO and protection of myocardial cells from injury.

ObjectiveTo investigate the unfavorable factors of intestinal mucosa repair after the intestinal epithelial injury in vivo in a mouse model of sepsis. MethodsThe method of cecal ligature and puncture (CLP) was used to induce sepsis and then the intestinal mucosa damage, epithelial cell apoptosis and the number of transformed goblet cells were observed, and the concentrations of serum TNF-αt, IL-1 and TGF-β1 and TFF3 ( trefoil factor 3) in small intestinal mucosa were determined. All above various laboratory examinations were made by different assays including H-E staining, western blot, ELISA and immunohistochemistry respectively. The experimental mice were divided into sepsis group and sham operation control group. The mice with sepsis were separately sacrificed 6 hours ( n = 7 ), 24 hours ( n = 7) and 48 hours ( n = 7) after CLP. Results In septic mice group, the injured intestinal mucosa was found 6 hours after CLP. The damage scores in mice 24 h and 48 h after CLP were higher than those 6 h after CLP, but there was no significant difference between those 24 h and 48 h after CLP. Moreover, a few goblet cells or other epithelial cells adjacent to the injured surface migrated onto the wound to cover the denuded area. The number of goblet cells was substantially decreased in mice of sepsis group 6 hours after CLP compared with sham operation control group. Compared with sham operation control group, levels of IL-1 and TNF-α significantly increased 3-4 times in mice of sepsis group at all intervals, and the phosphorylated caspase-3 increased 4 times. Although TFF3 assayed by using Western blot showed modest increase 6 h after CLP and it declined 24 h and 48 h later. A similar change was found in TGF-β1, it modestly increased 6h after CLP, but it didn't elevate 24 h and 48 h later. ConclusionsSevere sepsis keeps on the inflammatory reaction and epithelial cell apoptosis, preventing the repair of intestinal mucosa from injury.

Objective To investigate the protective effects of Ulinastatin on intestinal barrier damaged after cardiopulmonary resuscitation (CPR) in rats in order to illustrate the possible mechanism.Methods Twenty-one male SD rats were divided into three groups randomly (random number) including control group (sham group, n =7), cardiopulmonary resuscitation group (CPR group, n =7) and ulinastatin group (UTI group, n =7).The rats were anesthetized with pentobarbital sodium (45-60 mg/kg) by intraperitoneal injection.The rats of sham group were only treated with endotracheal intubation.Ulinastatin (100 000 U/kg) were injected via caudal vein 2 hours prior to CPR, and cardiac arrest was made in rats and cardiopulmonary resuscitation was carried out in the UTI group, while equivalent volume of sterile saline was used instead in the CPR group.Blood and ileum samples were obtained at 48 hour after restoration of spontaneous circulation (ROSC).The levels of tumor necrosis factor alpha (TNF-α) and interleukin-1β (IL-1β) were assayed by ELISA (enzyme-linked immunosorbent assay), the protein levels of caspase-3 were determined by western blot, the intestinal mucosa were stained by terminaldeoxynucleoitidyl transferase mediated nick end labeling (TUNEL) and ileac mucosa were observed under transmission electron microscope.Data were processed with SPSS 17.0 software.Results The plasma levels of TNF-α and IL-1β were dramatically higher in CPR group than those in other two groups (CPR vs.sham, P ＜ 0.01;CPR vs.UTI, P ＜ 0.05).Moreover, the tight junctions between cells obviously broadened and loosened in the CPR group were found under electron microscope, however, this phenomenon was not obvious in the UTI group.A large number of apoptotic cells were observed by TUNEL assay in the CPR group, but a small number of apoptotic cells were observed in the UTI group.The protein levels of caspase-3 in the UTI group were higher than those in sham group, but lower than those in CPR group (both P ＜ 0.05).Conclusions Ulinastatin has protective effects on the intestinal barrier damaged after cardiopulmonary resuscitation in rats by decreasing the proinflammatory mediators in the blood, reducing the expression of caspase-3and then reducing the numbers of apoptotic intestinal cells.

Objective To observe the effects of modified Jianpi Yishen Decoction on urinary osteopontin (OPN) in calcium oxalate nephrolithiasis patients after percutaneous nephrolithotomy (PCNL) or ureteroscope lithotomy (URL);To clarify the mechanism of modified Jianpi Yishen Decoction on the prevention of calcium oxalate kidney stones. Methods Totally 116 calcium oxalate nephrolithiasis patients were randomly divided into trial group (62 cases) and control group (54 cases). The trial group took modified Jianpi Yishen Decoction every other day, while the control group took potassium sodium hydrogen citrate granules three times a day. The concentrations of OPN, urinary calcium and urinary oxalic acid of the patients in the two groups were observed before treatment and 2 weeks and 4 weeks of treatment. Results The concentration of urinary OPN of 2 weeks and 4 weeks of the treatment in the trial group was significantly increased compared with before treatment (P<0.05), and the difference was statistically significant compared with the control group (P<0.05). The concentration of urinary OPN in the control group had no significant change after treatment (P>0.05). The differences in the concentrations of urinary calcium and urinary oxalic acid of the two groups between before and after treatment were not significant (P>0.05). Conclusion Modified Jianpi Yishen Decoction can effectively restrain the formation of the calcium oxalate stones by increasing the level of urinary OPN, which demonstrates effective prevention in the calcium oxalate nephrolithiasis patients after PCNL or URL.

Objective To evaluate the effects of diagnostic and treatment manipulation of the urinary diseases on serum prostate specific antigen(PSA) levels .Methods 80 male patients were recruited from urology surgery department of Shenzhen Tranditional Chinese Medicine (TCM ) Hospital ,Which included 13 cases with digital rectal examination (DRE) ,10 cases with catheterization , 12 cases with rigid cystoscopy ,17 cases with prostate biopsy ,28 cases with transurethral resection of the prostate (TURP) .Blood samples of 80 patients were collected before diagnostic and treatment manipulation of the urinary diseases and 24 h ,3 d ,7 d ,14 d af‐ter that ,respectively .Then ,serum total prostate antigen(TPSA) and free prostate antigen (FPSA) was measured .Results There was no effects of DRE on serum TPSA and FPSA levels(P>0 .05) .On the contrary ,serum TPSA and FPSA levels increased sig‐nificantly in patients with catheterization and cystoscopy(P<0 .05) ,and the duration was longer(7-14 d) .Serum TPSA and FPSA levels increased significantly(P<0 .05)in patients with TURP and biopsy at the 24th hour after manipulation and it began to de‐crease on the third day .Also ,the serum TPSA and FPSA levels decreased to baseline after 14 days .Conclusion There′re no effects of DRE on serum TPSA and FPSA levels .However ,serum TPSA and FPSA levels increase differently in patients with catheteriza‐tion ,cystoscopy ,biopsy and TURP ,but the durations were different ,too .

Objective Autophagy serves a role in the pathogenesis of chronic inflammatory diseases. The aim of the present study was to compare the autophagy levels of the peripheral blood mononuclear cells (PBMCs) in patients with chronic obstructive pulmonary disease (COPD) and healthy individuals and to assess the association between autophagy and the clinical parameters of COPD. Methods Samples of peripheral blood from 20 patients with stable COPD and 20 healthy controls were collected. PBMCs were harvested using Ficoll density gradient centrifugation. Levels of the autophagy-associated proteins ubiquitin-binding protein P62 (P62), microtubule-associated proteins 1A/1B light chain 3A (LC3 I/Ⅱ) and beclin-1 in PBMCs were detected by western blotting. Enzyme-linked immunosorbent assay kits were used to detect the serum concentrations of interleukin (IL)-6, IL-8 and tumor necrosis factor (TNF)-α. Results Western blotting demonstrated that the protein expression of P62 decreased (P62/GAPDH = 0.212 ± 0.089 vs. 0.378 ± 0.176,P = 0.001 1), but LC3 I/Ⅱ(4.94 ± 1.85 vs. 3.85 ± 1.06, P = 0.024 9) and beclin-1 (Beclin-1/GAPDH = 0.578 ± 0.126 vs. 0.149 ± 0.035, P<0.01) levels increased in patients with COPD compared with healthy controls. Serum levels of IL-6 [(11.96 ± 3.46) ng/L vs. (2.70 ± 1.72) ng/L, P < 0.01], IL-8[(20.38 ± 18.44) ng/L vs. (6.00 ± 4.08) ng/L, P=0.001 6] and TNF-α (29.50 ± 28.18 vs. 11.08 ± 12.07, P=0.010 6) increased in patients with COPD. The extent of PBMC autophagy was negatively correlated with FEV1% predicted, but positively correlated with levels of proinflammatory cytokines. The levels of autophagy in PBMCs in patients with COPD increased and were negatively correlated with FEV1% predicted and positively correlated with circulating levels of pro-inflammatory cytokines. Conclusions Autophagy may serve a role as a biomarker of the severity of COPD or as a therapeutic target for treatment of COPD.

OBJECTIVETo determine the optimal concentration of c-erbB2 antisense probe labeled with superparamagnetic iron oxide (SPIO) nanoparticles for in vivo tumor imaging in mice using magnetic resonance imaging (MRI).

METHODSThirty BALB/c mice bearing SK-Br-3 tumor were randomized into 5 groups to receive injections of different concentrations of SPIO-labeled c-erbB2 antisense probe (containing 6.0, 9.0, 12.0, 15.0, or 18.0 mg Fe/kg). MRI was performed before and 6 h after the injections, and the signal intensities of the tumor were compared among the groups. The tumor tissues were then dissected for microscopic examination with HE and Prussian blue staining.

RESULTSThe tumor-bearing mice all survived after injections of the probe at doses of 6.0, 9.0 and 12.0 mg, but injections at higher doses (15.0 and 18.0 mg) caused death in some mice. Injections of the probe at the doses of 12.0, 15.0 and 18.0 mg resulted in significant signal enhancement of the tumor (P<0.001) to allow visual identification, but the changes showed no significant differences among the 3 groups (P>0.05). Pathological examination revealed irregular structures of the tumor issue containing numerous heterogeneous tumor cells aligned into cancer nests; Prussian blue staining visualized scattered blue iron particles in the tumor issue, which was especially obvious in mice injected with 12.0, 15.0 and 18.0 mg labeled probe.

CONCLUSIONInjection of 12.0 mg/kg SPIO-labeled c-erbB2 antisense probe allows optimal tumor imaging in BALB/c mice using MRI.

Animals ; Antisense Elements (Genetics) ; Cell Line, Tumor ; Contrast Media ; Dextrans ; Genes, erbB-2 ; Magnetic Resonance Imaging ; Magnetite Nanoparticles ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Nanoparticles ; Nucleic Acid Probes ; genetics ; Xenograft Model Antitumor Assays

Objective:To explore the effects of protoporphyrin Ⅸ (PpⅨ)-mediated photodynamic therapy (PDT) on induction of apoptosis and death in colon cancer cell and the underlying mechanisms.Methods:The cell killing effect of PDT on HCT116 cell was determined by cell counting kit (CCK).The cells were divided into a control group,a single light group,a single PpⅨ group,and a PDT group.Hoechst 33342 and flow cytometry was used to assess the cell apoptosis.Western blot was employed to analyze the expressions ofbd-2,bax,and caspase-3.Reactive oxygen species (ROS) was detected by flow cytometry.Results:The viability of HCT116 cell was decreased gradually with the increase of irradiation dose (P<0.05).Compared to the other 3 groups,ROS production,the number of apoptotic cells and the protein expressions ofbax and caspase-3 in PDT group increased,while bcl-2 expression was decreased (P<0.05).Conclusion:PpⅨ-mediated PDT can enhance the apoptosis in HCT116 cell,which may be related to mitochondrial apoptosis pathway.

Objective To assess the performance of PET-CT in pulmonary carcinoid and to evaluate the efficacy of lymph node metastasis.Methods We retrospectively collected the data of 33 patients with primary pulmonary carcinoid in our center between November 2008 and March 2017.The results of PET-CT were analysed and the relationship among pathological sub-type, tumor size and SUVmax was compared.Results Thirty-three patients had a diagnosis of carcinoid tumor, typical in 10 patients and atypical in 23 patients.The major axis of the tumor was positively correlated with the SUVmax value (P<0.001), and the mean SUVmax of the atypical carcinoid was higher than that of the typical carcinoid(11.27 vs 3.54).The major axis of the tumor was controlled.One-way covariance analysis suggested that the pathological subtype was related to SUVmax(P=0.012).Overall, PET-CT had a sensitivity of 66.7％(95％CI: 12.5％-98.2％) and specificity of 90.9％(95％CI:83. 0％-95.5％).Conclusion Atypical carcinoids showed higher SUVmax than typical carcinoids, but the PET-CT perform-ance of lung carcinoids is not specific.There was a high specificity in evaluating lymph node metastasis.

OBJECTIVETo evaluate the clinical application of carbon nanoparticles labeled lymph node staining in curative laparoscopic resection for colorectal carcinoma.

METHODSSixty-five patients undergoing curative laparoscopic resection for colorectal carcinoma in the Sun Yat-sen Memorial Hospital between September 2011 and June 2013 were prospectively enrolled and randomly divided into label group (with carbon nanoparticles, n=34) and control group (without carbon nanoparticles, n=31). Association between labeled lymph nodes and metastasis was analyzed. The total number of retrieved lymph nodes and lymph nodes metastatic ratio were compared between the two groups.

RESULTSMean number of retrieved lymph node of the label group was higher as compared to the control group (22.3±4.2 vs. 15.4±3.5, P<0.05). The total number of retrieved lymph node was 725 in the label group and 478 in the control group. Among them, lymph node < 5 mm accounted for 4.6% (33/725) in the label group, which was higher than 2.0% (10/478) (P=0.025) in the control group. The number of black stain label lymph node was 412, with black stain ratio 56.8% (412/725) in the label group. Metastatic ratio of black stain nodes was significantly higher than that of non-stain nodes [28.6% (118/412) vs. 19.5% (61/313), P=0.005].

CONCLUSIONSThe technique of carbon nanoparticles labeled lymph node staining in curative laparoscopic resection for colorectal carcinoma is easy and effective, which can increase the retrieved number of lymph nodes, especially for nodes < 5 mm. The black stain lymph nodes indicate higher risk of metastasis.

Carbon ; Colorectal Neoplasms ; Humans ; Laparoscopy ; Lymph Node Excision ; Lymph Nodes ; Lymphatic Metastasis ; Nanoparticles ; Staining and Labeling