AIM:Sodium butyrate has antitumor effects on colon cancer cells such as inhibiting cell growth and promoting differentiation and apoptosis.The aim of this study is to investigate whether sodium butyrate induces apoptosis in human colon cancer cell line HT-29 and to examine the intracellular mechanisms involved,especially the role of caspase activation in the process.METHODS:HT-29 cells were cultured to logarithmic phase before treatment with sodium butyrate at concentration of 5.0 mmol/L and caspase inhibitors at the concentration of 20 ?mol/L.The latter were added in the medium ahead of sodium butyrate for 1 h.Then,the staining of Annexin V-FITC and PI were used to analyze HT-29 apoptosis and the dye JC-1 was applied to detect mitochondrial membrane potential by flow cytometry.Caspase activity within the cells was measured respectively using a specific caspase activity assay kit and a microplate reader.RESULTS:Preincubation of HT-29 cells with sodium butyrate significantly increased apoptosis [(35.40?0.70)%] and decreased mitochondrial membrane potential(5.53?0.91).This effect was blocked when pretreatments were enforced with z-VAD-fmk,z-DEVD-fmk and z-LEHD-fmk.The apoptosis percentages were(1.33?0.59)%,(1.40?0.53)% and(1.27?0.91)%,respectively and mitochondrial membrane potentials were 9.80?1.15,10.23?0.50 and 10.33?1.02,respectively.However,the role of reduction by z-IETD-fmk,which presented the apoptosis percentage of(32.10?2.33)% and mitochondrial membrane potential of 5.93?1.31,was not observed.An enhancement of caspase-3 and-9 activities(2-3-fold)but no change of caspase-8 activity was confirmed.CONCLUSION:Apoptosis of HT-29 colon carcinoma cells induced by sodium butyrate is tightly linked to caspase activation via mitochondrial pathway other than tumor necrosis factor-alpha and has the potential to inhibit proliferation and thereby may contribute to the progression of colon cancer.

Animals ; Biomedical Research ; Humans ; Interdisciplinary Communication ; Mental Disorders ; Nervous System Diseases ; Research Design ; trends

Objective To investigate the effects of motilin on the voltage dependent potassium channel and L-type calcium channel currents in rat proximal colon smooth muscle cells (PCSM) and to explore its mechanism in increasing colonic motility.Methods PCSM were isolated by collagenase.The voltage dependent potassium channel transit outward current (IKA ) and delayed rectifier current (IKdr) and L-type calcium currents (ICa(L)) were measured by whole cell patch clamp technique.Groups were analyzed by paired t-test.Results There was no significant effect of motilin on IKA and IKdr.L-type calcium channel was dose-dependently activated by motilin from 0.5 × 105 mmol/L to 10.0 ×10-5 mmol/L.At 6 × 10-5 mmol/L motilin and under - 10,0 and 10 mV stimulating voltage,maximum current density increased by 154.61％,62.69％ and 21.02％ respectively and activation kinetics curve obviously left shifted.Half activation voltage decreased from (2.740±1.211) mV prior administration to ( - 25.290 ± 0.614) mV (t =8.534,P =0.007 ) and there was no significant difference in slope factor. Conclusions Motilin increases colonic smooth muscle contraction by promoting calcium influx. However the frequency of colonic smooth muscle contraction could not change with frequency of equilibrium potential and action potential of colonic smooth muscle.

[ ABSTRACT] AIM:To investigate the potential role of exogenous hydrogen sulfide ( H2 S) and ATP-sensitive po-tassium ( KATP ) channels in chronic stress-induced colonic hypermotility.METHODS:Male Wistar rats were divided into water avoidance stress ( WAS) group and sham WAS ( SWAS) group.Organ bath recordings were used to test the contrac-tile activity of colonic strips.The effects of H2 S donor NaHS and pretreatment with glibenclamide on the contractions of co-lonic smooth muscle were studied and the IC50 of NaHS was calculated.The localization and expression of the subunits of KATP channels were determined by the methods of immunohistochemistry and Western blotting.RESULTS:WAS increased contractile activity of colonic strips.NaHS concentration-dependently inhibited the spontaneous contractions of strips from the SWAS and WAS rats.The IC50 of NaHS for longitudinal muscle ( LM) and circular muscle ( CM) of the WAS rats was 0.2033 mmol/L and 0.1438 mmol/L, significantly lower than those of the SWAS rats.Glibenclamide significantly in-creased the IC50 of NaHS for LM and CM from the SWAS and WAS rats.In both SWAS and WAS rat colon, Kir6.1, Kir6.2 and SUR2B were expressed on the plasma membrane of the smooth muscle cells.WAS treatment resulted in up-reg-ulation of the expression of Kir6.1 and SUR2B in the colon devoid of mucosa and submucosa.CONCLUSION: The in-creased expression of Kir 6.1 and SUR2B in colonic smooth muscle cells may be a defensive response to chronic WAS.H2 S donors may have potential clinical effect on treating chronic stress-induced colonic hypermotility.

Objective To investigate etiology and clinical features of acute pancreatitis (AP) in young patients. Methods In total, 423 patients with AP admitted to Renmin Hospital of Wuhan University, Wuhan, Hubei province during January 2006 to December 2009 were retrospectively analyzed, 55 cases aged 34 years or less as young group and another 55 cases aged more than 34 years as middle- and elderly-age group. Their etiology, clinical features and illness scores based on some standard criteria were assessed. Results Proportion of varied etiologies of AP in young group was different from that in middle- and elderly-age one (x2 = 17. 107, P = 0. 009) , mainly as biliary and idiopathic pancreatitis in young group and as biliary and alcoholic pancreatitis in middle- and elderly-age one. Incidence of diet-related and idiopathic pancreatitis were higher in young group than that in middle- and elderly-age one ( x2 =4. 853, P = 0. 028 and x2=4. 274, P = 0. 039 ). Scores of the acute physiology and chronic health evaluation II ( APACHE II ) , Ranson criteria and the bedside index for severity in acute pancreatitis ( BISAP) were all lower in young group than those in middle- and elderly-age one (t = 5. 381 , P = 0. 000; t = 4. 388 , P = 0. 000 and t=3. 083, P = 0. 003 ) . There was no statistically significant difference in scores of computed tomography severity index ( CTSI) between young group and middle- and elderly-age one (t = ±0. 750, P > 0. 05 ) . Recurrence rate of AP was lower in young group than that in middle- and elderly-age one ( x2 =4. 251, P = 0. 039). Cholecystitis was more complicated in young group, pregnancy was more complicated in young group (x2 =4. 151 , P =0. 042) , and elevated blood glucose was more complicated in middle- and elderly-age one (x2 = 13. 285 , P = 0. 000 ) . Conclusions Etiology of AP varies in young patients with more dietary factors, occurrence of systemic complications and risks for death are lower in young group than those in middle- and elderly-age one, and there was no statistically significant difference in local complications and severity of illness between the two groups.

Objective To observe the clinical efficacy of infrared thermal therapy against pseudomonas ae-ruginosa infection on deep partial-thickness burn wound. Methods Forty-three patients in our hospital with main-ly deep partial-thickness burn wound from January 2015 to October 2016 were randomly enrolled to the treatment group(TG,treated with sulfadiazine silver + infrared thermal therapy)and the control group(CG,treated with sulfadiazine silver only). Scores of wound exudation,positive rate of pseudomonas aeruginosa on wound,wound healing rate,wound healing time and overall evaluation of wound healing on the day of 0,3,7,14,21,28 after treatment were conventionally recorded. Adverse effects in TG and CG were also observed. Results (1)Age,sex and burn surface area of patients were found no statistically significant difference between the two groups(P>0.05). (2)On the day of 3,7 and 14,wound exudation score of TG was significantly lower than that of CG(P<0.05);On the day of0,21 and 28,wound exudation score of the two groups were almost same.(3)Positive rate of pseudomonas aeruginosa between the two groups on the day of 7,14 and 21,TG was significantly lower than CG(P < 0.05). (4)Wound healing rate of TG on the day of 7,14 and 21 was higher than CG,which was statistically significant difference in the 2 groups(P<0.05);Wound healing time of TG patients[(21.1 ± 6.5)day]was significantly shorter than that of CG patients[(26.2 ± 6.5)day](P<0.05).(5)Overall evaluation of wound healing of TG was better than that of CG on the day 14 and 21(P<0.05). Conclusions Infrared thermal therapy could reduce the secretion of deep partial-thickness burn wound and effectively control pseudomonas aeruginosa infection. Furthermore,infrared thermal therapy finally improved wound healing rate and shortenedwound healing time of burn wound.

Aim To investigate the effects of emodin on the contraction of gallbladder smooth muscle(GBSM)and the L-type calcium current in GBSM cells.Methods Gallbladder muscle strips were obtained from adult guinea pigs and the resting tension was recorded.Gallbladder smooth muscle cells were isolated by enzymatic digestion,and calcium current was recorded by the whole-cell patch clamp method.Results Emodin-induced contraction of GBSM was significantly attenuated by pretreatment with nifedipine.Emodin increased the L-type calcium current in a dose-dependent manner.When 10 ?mol?L-1 emodin was applied to GBSM cells,the amplitude of L-type calcium current at +10 mV was enhanced by(45.2?2.26)%.In the presence of PKC inhibitor,staurosporine,emodin did not significantly affect the calcium current.Conclusion Emodin enhances L-type calcium current via PKC-dependent pathway and promotes gallbladder contraction.

OBJECTIVETo establish the steriod-induced osteoporosis model with the type of Kidney-Yin deficiency.

METHODSTotally 45 female Kunming mice were randomly divided into normal group,model group and Liuwei Dihuang pills(Chinese character: see text)group. The model was established by intramuscular injecting of Dexamethasone. Liuwei Dihuang pills (Chinese character: see text) group was administered orally with Liuwei Dihuang pills (Chinese character: see text). The signs and symptoms of mice were observed dynamically. All the animals were sacrificed at the end of the 6th weeks. The level of ACTH, cAMP, cGMP, TSH and E2 in serum were detected to evaluate deficiency of Kidney-Yin. Morphological changes and bone density were observed to evaluate osteoporosis.

RESULTS(1) Compared with control group, mice in model group appeared obvious Kidney-Yin deficiency symptoms, including hair dry, restlessness, excitability, hard stool, and yellow. (2) Compared with control group,the weight of mice in model group gained slower (P<0.01); the index of adrenal gland,liver and spleen decreased (P<0.01, P<0.01 ,P<0.01); the level of ACTH and TSH increased (P<0.01 ,P<0.01), the level of E2 decreased (P<0.01) and the ratio of cAMP/cGMP increased (P< 0.05). (3)Compared with control group,the bone density of lumbar vertebra and femur in model group were significantly decreased (P<0.01, P<0.05); HE staining revealed osteoporosis in model group mice. (4)However, the Liuwei Dihuang pills (Chinese character: see text) group can partly antagonize the inhibition of the HPA axis, alter the disordered sex hormone and the ratio of cAMP/cGMP, and reverse the osteoporosis partly.

CONCLUSIONthe model of osteoporosis with type of Kidney-Yin deficiency could be established by Dexamethasone intramuscular injection. With less interference, it wight be a stable and reliable modeling method for integration of disease and syndrome in TCM.

Animals ; Bone Density ; Dexamethasone ; toxicity ; Disease Models, Animal ; Female ; Kidney Diseases ; etiology ; Medicine, Chinese Traditional ; Mice ; Osteoporosis ; chemically induced ; Yin Deficiency ; complications

Objective:It is demostrated that the porous protein-mineral mechanics model could provide more accurate prediction for biomaterial properties of dentine compared with the other established models.This paper would use the model to reevaluate the mechanical properties and its interacting mechanism of human dentine.Method:By using a porous proteinmineral mechanics model,the effect from the interactions between tubules,pedtubular and intertubular matrix on dentine microstructure was discussed.Results:The dentinal micromechanical properties were dependent on the tubular direction,and the absolute values of the stresses derived from the hydraulic and gas tubular pressures increased parabolically with theincreasing diameter of the tubules.It was also found that the effective elastic constants of the dentine microstructure would vary with the aging and the distribution of mineral and collagen within peritubular and intertubular matrix of detine.Conclusions:The theoretical analyses provided in this paper demonstrated that the microstructural characteristics of tubules,peritubular and intertubular dentinal matdx could have different influences on the micromechanical properties of human dentine,which showed the validity of porous protein-mineral mechanics model,and the limitation of some models that neglected the interacting mechanism.

Objective:It is demostrated that the porous protein-mineral mechanics model could provide more accurate prediction for biomaterial properties of dentine compared with the other established models.This paper would use the model to reevaluate the mechanical properties and its interacting mechanism of human dentine.Method:By using a porous proteinmineral mechanics model,the effect from the interactions between tubules,pedtubular and intertubular matrix on dentine microstructure was discussed.Results:The dentinal micromechanical properties were dependent on the tubular direction,and the absolute values of the stresses derived from the hydraulic and gas tubular pressures increased parabolically with theincreasing diameter of the tubules.It was also found that the effective elastic constants of the dentine microstructure would vary with the aging and the distribution of mineral and collagen within peritubular and intertubular matrix of detine.Conclusions:The theoretical analyses provided in this paper demonstrated that the microstructural characteristics of tubules,peritubular and intertubular dentinal matdx could have different influences on the micromechanical properties of human dentine,which showed the validity of porous protein-mineral mechanics model,and the limitation of some models that neglected the interacting mechanism.