[ ABSTRACT] AIM: To investigate the protective effect of mesenchymal stem cell ( MSC)-conditioned medium (MSCCM) on myocardial cell line H9c2 and its mechanism.METHODS:Verification of MSC was performed by flow cy-tometry analysis, followed by MTT assay to determine the optimal incubation time of MSCCM with myocardial cells.The cells were divided into 4 groups:normal ( N) group, model ( M) group, M+MSCCM group and MSCCM group.The cells in M+MSCCM group and MSCCM group were pre-incubated with MSCCM for 24 h.The cells in M group and M+MSCCM group were treated with 300 μmol/L H2 O2 for 4 h to imitate oxidative injury of myocardial cells.Mitochondrial membrane potential and apoptotic rate of injured myocardial cells were detected by flow cytometry.The ROS production was measured by fluorescence microscopy.The nuclear translocation of Nrf2 and expression of HO-1 was examined by Western blot.RE-SULTS:No difference of mitochondrial membrane potential, apoptotic rate or ROS production between MSCCM group and N group was observed (P>0.05).The mitochondrial membrane potential depolarization, apoptotic rate and ROS produc-tion in M+MSCCM group were significantly lower than those in M group ( P<0.01 ) .The nuclear translocation of Nrf2 and expression of HO-1 in the myocardial cells were increased with MSCCM incubation time prolonged.CONCLUSION:MSCCM protects the myocardial cells against oxidative injury induced by H2 O2 .The anti-oxidative mechanism would be as-sociated with the activation of Nrf2/ARE pathway.

Aim To investigate the protective effect of isorhamnetin on H9 C2 myocardial cell line and its mechanisms. Methods The toxicity and optimal pro-tective concentration of isorhamnetin were determined by MTT assay. The experimental subjects were divided into four groups:group N ( normal ) , group M ( mod-el) , group M + ISO ( model + isorhamnetin ) , and group ISO ( isorhamnetin only ) . Group M +ISO and ISO were pre-incubated with isorhamnetin for 12 hours while other groups with plain DMEM. Group M and M+ ISO were treated with 300μmol · L-1 H2 O2 for 4 hours after pre-incubation. Mitochondrial membrane potential depolarization of H9 C2 was measured by fluo-rescence microscope. Apoptotic rate and ROS produc-tion of injured myocardial cell line were detected using
flow cytometry. The oxidative indictors were measured by spectrophotometry. The expressions of cytoplasmic cytochrome C, caspase-9, caspase-3, Bcl-2, Bax, Nrf2 and HO-1 were examined by Western blot. Result There was no difference in mitochondrial membrane potential depolarization, apoptotic rate, ROS produc-tion, oxidative indictors production and expressions of cytoplasmic cytochrome C, caspase-9,caspase-3, Bcl-2 , Bax between groups ISO and N ( P>0. 05 ) . Apop-totic rate, ROS production, expressions of cytoplasmic cytochrome C, caspase-9, caspase-3, Bax, MDA pro-duction of group M+ISO were significantly lower than those of group M ( P < 0. 01 ) . And mitochondrial membrane potential, Bcl-2, CAT, SOD and GSH-Px of group M + ISO were increased compared to group M .
Nuclear translocation of Nrf2 and expression of HO-1 in myocardial cell line were increased with the prolonged isorhamnetin incubation time. Conclusion Isorham-netin could protect myocardial cell line against H2 O2-induced oxidative injury and apoptosis through the in-
terruption of mitochondrial dependent apoptotic path-way and activation of Nrf2/ARE pathway.

Aged ; China ; Confusion ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Medicine in Literature

Objective To investigate the effect of continuous blood purification (CBP) on pro-or anti-inflammatory immune function and prognosis in patients with sepsis. Methods One hundred and two patients with sepsis were randomized into the CBP group (60 cases) and the control group (42 cases). The patients in the control group were treated with conventional therapy, and the patients in the CBP group received at least 72 h CBP treatment additionally. The APACHEⅡ score, SOFA score, the 28 day survival rate and ICU length of stay were recorded and levels of spleen Th1, Th2 were assessed by FACS flow cytometry. Enzyme linked immunoadsordent assay (ELISA) was used to determine the levels of serum IL-1, IL-10 and TNF-α before and at 24, 48, 72 h after the treatment. Results The APACHEⅡscore and SOFA score decreased markedly in the CBP group after the treatment (Р <0.05). The period of staying in ICU of patients in the CBP group was shorter than that of patients in the control group (Р < 0.05). There was no significant difference of the 28 day survival rate between the two groups (91.6% vs 71.2%, Р > 0.05). Compared with the control group, levels of IL-1, IL-10 and TNF-α were decreased markedly, and the ratio of Th1 / Th2 was increased significantly at 72 h after the treatment in the CBP group (Р < 0.05). Conclusion CBP can eliminate inflammatory mediators, and help to enhance the immune function, and restore the balance of Th1 / Th2 in patients with severe sepsis.

Age Determination by Skeleton ; Body Height ; drug effects ; Child ; Child Development ; drug effects ; Drug Therapy, Combination ; methods ; Female ; Gonadotropin-Releasing Hormone ; administration & dosage ; analogs & derivatives ; therapeutic use ; Growth Hormone ; administration & dosage ; therapeutic use ; Humans ; Menarche ; Puberty, Precocious ; diagnosis ; drug therapy ; physiopathology ; Treatment Outcome

Objective To study the left atrial appendage (LAA) function and haemodynamics by transesophageal echocardiography,and predict high risk of thrombosis.Methods Eighty-nine consecutive patients were studied using transesophageal echocardiography.According to with spontaneous echo contrast (SEC) or not,all patients were divided into SEC group (n =17) and non-SEC group (n-=72).LAA emptying peak flow velocity (LAA-EV),filling peak flow velocity (LAA-FV) were measured by pulsed Doppler.The middle segment of the LAA medial wall systolic peak velocity (LAA-TSV) and diastolic peak velocity (LAA-TDV) were obtained by tissue Doppler imaging.LAA color Doppler and M-mode color Doppler images were acquired and derived the LAA emptying flow propagation velocity (LAA-EPV).Results LAA-EV,LAA-FV,LAA-TSV,LAA-TDV,LAA-EPV of SEC group were lower than those of non-SEC group (P ＜0.05).LAA M-mode color Doppler had four characteristic patterns,N pattern (normal pattern) which flow eject from LAA apical to LAA orifice,R pattern (reverse pattern) which flow eject from LAA orifice to LAA apical,V pattern (vertical pattern) vertical signals seen in systole and without efficient flow,S pattern (stasis pattern) almost no flow signal.V,R and S patterns may indicate high risk of SEC and thrombosis.LAA-EPV had strong correlation with LAA-EV (r =0.742,P ＜0.001),and moderate correlation with LAA-TSV (r =0.462,P ＜0.01),LAA-EV had strong correlation with LAA-TSV (r =0.600,P ＜0.01).Conclusions LAA dysfunction would result in hypokinetic LAA wall motion and reduced blood flow velocity.Emptying abnormalities of LAA might be the important reasons for blood stasis and thrombus formation.Emptying characteristic low patterns of V,R and S might be novel methods to predict high risk of thrombus.

Objective To investigate the effects of Xuebijing on the regulations of tumor necrosis factor-α(TNF-α) 、von Wilebrand factor(vWF)、interleukin-6 (IL-6) and the treatment effectiveness in pulmonary contusion patients.Methods Fifty-one pulmonary contusion patients admitted into the hospital from January 2010 to January 2012 were enrolled in the study and were randomly divided into two groups (treatment group and control group).The levels of the TNF-α、vWF 、IL-6 in the pulmonary contusion patients were measured by enzyme linked immunosorbent assay to investigate the change of the inflammatory cytokines and the treatment effectiveness in treatment group and control group.Results The hospitalization time of treatment group and control group was (19.83 ± 2.35) d,(21.52 ± 2.73) d respectively,the hospitalization time of the treatment group was significantly shorter than the control group,the difference was statistically significant (t =2.38,P ＜0.05) ;The incidence of ARDS was 14.81％ and 41.67％ respectively,the incidence of ARDS of the treatment group was significantly lower,the difference was statistically significant (x2 =4.6,P ＜ 0.05) ;The inflammatory mediators index of the two groups showed as follws:the first day of the TNF-α(181.35 ± 14.25) ng/L,(179.17 ± 16.28) ng/L,the third day of the TNF-α(169.21 ± 16.42) ng/L,(179.18 ± 17.31) ng/L,the fifth day of the TNF-α(131.78 ± 13.02) ng/L,(140.84 ± 14.27) ng/L,the seventh day of the TNF-α (99.45 ± 16.02)ng/L,(110.56 ± 17.09) ng/L;The first day of the vWF (178.73 ± 22.09) ng/L,(177.73 ± 23.02) ng/L,the third day of the vWF (208.46 ± 14.13) ng/L,(220.53 ± 23.02) ng/L,the fifth day of the vWF (252.32 ±15.22) ng/L,(266.40 ±25.79) ng/L,the seventh day of the vWF(201.49 ±9.21) ng/L,(217.27 ±22.68)ng/L;The first day of the IL-6 (172.14 ± 12.41) ng/L,(169.53 ± 13.55) ng/L,the third day of the IL-6 (147.24 ± 16.33) ng/L,(158.41 ±21.04) ng/L,the fifth day of the IL-6(126.36 ± 11.37) ng/L,(137.21 ± 21.23) ng/L,the seventh day of the IL-6 (114.03 ± 10.26) ng/L,(122.16 ± 13.59) ng/L,the differences within the group and between the two groups were significant(within the group:F =206.58,218.57,198.35 respectively,P ＜ 0.01 ;between the groups:F =19.053,17.259,17.235 respectively,P ＜ 0.01).And in the third,fifth and seventh day,3 indicators of treatment groups were lower than the control group (P ＜ 0.01).Conclusion Xuebijing can inhibit inflammatory transmitters,relieve inflammatory reactions,decrease the rate of ARDS incidence,shorten the hospitalization time and help to improve the prognosis of the patients with acute pulmonary contusion.

Objective To explore the feasibility,advantages and disadvantages of combined trans- gastric and transcolonic routes for NOTES.Methods A female swine was used in this study.Transgastric entrance was the first,followed by transeolonie entrance.A dual-channel endoscope was inserted through the porcine mouth into the gastric cavity and penetrated into the peritoneal cavity through the puncture and bal- loon dilatation of the gastric wall.Then under direct visualization through the transgastric approach,the other endoseope was advanced into the peritoneum.Using the two endoscopes inside the peritoneal cavity,collabo- rative peritoneoseopy was performed by the two endoscopists.After the examination the incisions in the stom- ach and the colon wall were closed with Endoclips.The animal was sacrificed for post-mortem examination with particular attention to the entrance sites and presence of any complications related to the access or to ma- nipulations inside the peritoneal cavity.Results No hemorrhage oecurred during the puneture and balloon dilatation or bow-knife cutting of the gastric wall or the eolonic wall.The liver was damaged while a needle knife penetrated the gastric wall.On the contrary,no organs were damaged during the needle knife penetra- ted the eolonic wall under direet visualization through the transgastric approach.It was difficult to find the gallbladder or the oviduct with a"single arm",but it was easy to see them with the double routes.It was easier to close the colonic incision than to close the gastrie wall with Endoclips.Conclusion Combined transgastric and transcolonic route for NOTES is feasible and it seems to be easier to show a target compared with a single route.

OBJECTIVETo observe the effects of Chailing Decoction (CD) on transforming growth factor-beta1 (TGF-beta1), connective tissue growth factor (CTGF), renal cell apoptosis and proliferation in rats with chronic cyclosporine A nephropathy (CCN), and to explore its possible mechanism for inhibiting renal fibrosis.

METHODSThe CCN rat model was prepared using oral administration of cyclosporine A (CsA, 30 mg x kg(-1) x d(-1)). Meanwhile, they were treated with CD (3 g x kg(-1) x d(-1)) by gastrogavage. The serum blood urea nitrogen (BUN), serum creatinine (SCr), and creatinine clearance rate (CCr) were measured by the end of the fourth week of the experiment. The kidneys were taken out on the next day. The degree of renal fibrosis was detected using Masson staining. The protein and gene expressions of TGF-beta1, and CTGF were observed using immunohistochemical assay and RT-PCR. The renal cell apoptosis rate and the proliferation index were detected by flow cytometry.

RESULTSCompared with the control group (BUN: 6.123 +/- 0.588 mmol/L; SCr: 75.654 +/- 8.196 micromol/L; CCr: 0.539 +/- 0.169 mL/min), the renal function of the model group (BUN: 11.600 +/- 1.437 mmol/L; SCr: 101.985 +/- 10.809 micromol/L; CCr: 0.272 +/- 0.060 mL/min) obviously declined (P < 0.01). The collagen deposition in the renal interstitial area significantly increased. The protein and mRNA expressions of TGF-beta1, and CTGF in the tubular epithelial cells and the mesenchymal cells were significantly enhanced (P < 0.01). The cell proliferation index and the apoptosis rate both increased, but the ratio of apoptosis to proliferation (0.317 +/- 0.059) decreased more than that in the control group (0.680 +/- 0.150, P < 0.01). After treatment by CD, the renal function (BUN: 7.340 +/- 0.857; SCr: 84.923 +/- 10.627; CCr: 0.405 +/- 0.081) was significantly enhanced (P < 0.05, P < 0.01), the collagen deposition decreased, the high protein and mRNA expressions of TGF-beta1 and CTGF were down-regulated (P < 0.01), the ratio of apoptosis to proliferation increased (0.650 +/- 0.092, P<0. 01).

CONCLUSIONCD could improve the renal function of CCN model rats, inhibit the expressions of TGF-beta1 and CTGF, and recover the balance between the renal cell apoptosis and proliferation by inducing cell apoptosis and inhibiting cell proliferation, thus delaying the renal fibrosis process.

Animals ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Connective Tissue Growth Factor ; metabolism ; Cyclosporine ; adverse effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Fibrosis ; Kidney ; drug effects ; metabolism ; pathology ; Kidney Diseases ; chemically induced ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism

Objective To investigate the protective effects of recombinant human erythropoietin (rHuEPO) on caecal ligation and puncture (CLP)-induced acute kidney injury (AKI).Methods A total of 260 healthy male Sprague-Dawley rats (250-300 g) were randomly divided into 6 groups:normal control group,sham group,CLP model group,the large dose rHuEPO (5000 U/kg)group,the middle dose rHuEPO (1000 U/kg) group,and the small dose rHuEPO (500 U/kg) group.The rat models of sepsis were established by CLP.In treatment groups,rats were treated with rHuEPO through caudalis injection after CLP surgery.Each group was divided into 2-,6-,12-,24-,36-hour subgroups with 10 rats.Rats were sacrificed and the tissue samples including kidney and blood samples were collected.The kidney function,plasma cytokines [interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α)],kidney injury moleclue 1 (KIM-1) and inducible nitric oxide synthase (iNOS)were measured.Cytokines were determined by ELISA method.The expression of nuclear factor-kappaB (NF-κB) protein in kidneys were detected by immnunohistochemistry method.Pathological changes of kidney tissues were observed by light and transmission electron microscopy for cytokine content and apoptosis.Results Compared with CLP model group,renal function,the levels of TNF-α,IL-6,KIM-1 and iNOS in serum,the expression of NF-κB,significantly decresed in large dose rHuEPO group (all P ＜ 0.05).rHuEPO also lessened the histological changes in large dose group.rHuEPO did not lessen the histological changes in others.Conclusion rHuEPO can inhibit the levels of TNF-α,IL-6 and iNOS in serum,thus modify the inflammatory response and provide protective effects against acute kidney injury induced by sepsis.