Aim To clarify the role of autophagy in the regulation mechanism of epithelial-mesenchymal transition(EMT) in hepatocytes and the transition to myofibroblast. Methods BNL CL. 2, a mouse embryonic hepatocyte, and BNL CL. 2 cells knocked-down BECN1 genes were treated with TGF-β1(2 μg . L-1) and curcumin(10, 20, 30 jimol • L - 1 ) . Real-time PCR and Western blot were used to detect Beclin-1, LC3, Vimentin and α-SMA gene and protein expression in BNL cl. 2 cells. The optical microscope was used to observe the changes of cell morphology. And the changes in cell autophagy were detected by GFPLC3 plasmid transfection. Results After the intervention of TGF-β1, the expression of Vimentin and α-SMA, the mesenchymal marker, increased and the autophagy decreased. Curcumin could inhibit Vimentin and a-SMA expression in hepatocytes. Moreover, curcumin could increase the autophagy related genes and protein expression of Beclin-1 and LC3 in hepatocytes. Curcumin could increase the level of autophagy and inhibit the cell transformation to myofibroblast-like cells. After interfering with BECN1, the effect of curcumin inhibiting hepatic EMT was partially blocked. Conclusions Curcumin could inhibit the process of EMT of hepatocytes by increasing the level of autophagy. The occurrence and development of liver fibrosis was consequently inhibited.

OBJECTIVETo study the color reappearance effect of metal-ceram restoration and foundry-ceram restoration using Crystaleye spectrophotometer.

METHODS58 metal-ceram restorations and 58 foundry-ceram restorations according to the result of the Crystaleye spectrophotometer were made respectively. The deltaE between restorations and natural teeth as referenced were analyzed. And satisfaction of dentists and patients were evaluated.

RESULTSThe deltaE between metal-ceram restorations and natural teeth was 7.13 +/- 0.74. The deltaE between foundry-ceram restorations and teeth was 1.47 +/- 0.84. There were statistical differences between the deltaE (P < 0.01). The satisnatural faction of dentists and patients to the foundry-ceram restorations were higher than to the metal-ceram restorations (P < 0.05).

CONCLUSIONCrystaleye spectrophotometer can provide accurate reference for foundry-ceram restoration, but for metal-ceram restoration it is not accurate.

Color ; Humans ; Metals ; Spectrophotometry ; Tooth

OBJECTIVEThis study aimed to explore the scene-trait coping style of military rescuers in Wenchuan earthquake in an effort to provide scientific evidences for mental intervention program for Chinese military personnel.

METHODSBy cluster sampling, a total of 151 military rescuers and 331 control servicemen were administered the military personnel scene-trait coping style scale (MPSTCSS).

RESULTSAll active coping factor scores, and passive coping factor scores of affection, health and economy in the rescuer group were significantly higher than those in control group (P < 0.05). The 21-above age subgroup, the 3-year plus service subgroup, and the officer subgroup had significantly higher active coping factor scores on military tasks, military experience and personal development than those of the 21-below age subgroup, 3-year minus service subgroup and the soldier subgroup, respectively (P < 0.05 or P < 0.01).

CONCLUSIONThe earthquake relief servicemen can cope with stressful situations better than control group by taking active coping style. The officers, servicemen older than 21 years, and servicemen with more service duration than 3 years could usually take active coping style.

Adaptation, Psychological ; Disasters ; Earthquakes ; Humans ; Military Personnel ; psychology ; Stress, Psychological

AIMTo study whether store-operated Ca2+ channel (SOC) is present in rat colonic smooth muscle cells.

METHODSIntracellular Ca2+ ([Ca2+]i) changes induced by thapsigargin- or caffeine-activated SOC channel were measured in enzymatically dissociated rat colonic smooth muscle cells with the fluorescent indicator Fura-2/AM.

RESULTSIn the absence of external Ca2+ , the sarco-endoplasmic reticulum Ca2+ pump inhibitor thapsigargin (1 micromol/L) and ryanodine receptor (RyR) activator caffeine both transiently elevated [Ca2+]i from (68.32 +/- 3.43) nmol/L to (240.85 +/- 12.65 ) nmol/L, (481.25 +/- 34.77) nmol/L. A subsequent reintroduction of Ca2+ into the extracellular solution resulted in [Ca2+]i further elevated to (457.55 +/- 19.80) nmol/L, (1005.93 +/- 54.62) nmol/L; (643.88 +/- 34.65) nmol/L, (920.16 +/- 43.25) nmol/L, respectively. And the elevated response was blocked by lanthanum (1 mmol/L), but was insensitive to L-type voltage calcium channels blocker verapamil and membrane depolarization.

CONCLUSIONSOC activated by store depletion are present in rat colonic smooth muscle cells. And we further prove the existence of such Ca2+ channels in excitable cells.

Animals ; Caffeine ; pharmacology ; Calcium ; metabolism ; Calcium Channels ; physiology ; Colon ; cytology ; Fura-2 ; metabolism ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Rats ; Rats, Wistar ; Thapsigargin ; pharmacology

OBJECTIVETo determine the optimal thread pitch for an experimental cylinder implant in Ansys Work-bench Design Xplorer environment.

METHODSFinite element models of segment jaw bone with a V-shaped thread implant were created. The thread pitch (P) was set from 0.5 mm to 1.6 mm. The maximum Equivalent stresses (EQV stresses) in jaw bone and in implant were evaluated.

RESULTSUnder axial load, the amplification of maximum EQV stresses in cortical bone, cancellous bone and implant were 7.1%, 123.4% and 28.7% respectively. Under bucco-lingual load, the amplification of maximum EQV stresses in cortical bone, cancellous bone and implant were 2.8%, 28.8% and 14.9% respectively. When P exceeded 0.8 mm, the response curve curvature of maximum EQV stresses in jaw bone and in implant to P was ranged from -1 to 1.

CONCLUSIONStresses in cancellous bone are more sensitive to thread pitch than in cortical bone. Stresses in jaw bone under axial load are easier affected by thread pitch than under bucco-lingual load. Thread pitch plays a greater role in protecting dental implant under axial load than under bucco-lingual load. Thread pitch exceed 0.8 mm should be the optimal design in a cylinder implant, but oversized pitch should be avoided too.

Biomechanical Phenomena ; Computer Simulation ; Dental Implants ; Dental Prosthesis Design ; Finite Element Analysis ; Mandible ; Stress, Mechanical

OBJECTIVETo investigate the infection status and epidemiological characteristics of influenza virus and respiratory syncytial virus (RSV) in influenza-like illness (ILI) of children ( ≤ 14 years) in Wuhan area from 2008 to 2012.

METHODSA total of 2854 cases of ILI patients ( ≤ 14 years) in a hospital of Wuhan were recruited in the study from July 2008 to June 2012. The sample of pharyngeal swab was collected from each patient, to extract the virus nucleic acids. Real-time fluorescent quantitation reverse transcription PCR (RT-PCR) method was applied to detect the subtypes of influenza virus and RSV, and then analyzed the time and age characteristics.

RESULTSOut of the 2854 cases, 758 (26.6%) were positive for influenza virus,including 547 (19.2%) influenza A virus positive samples and 211 (7.4%) influenza B virus positive samples. Usually, there were two peaks present in the annual curve of influenza virus, namely summer peak and winter/spring peak. The positive rate of influenza virus in 6-14 years old children (48.0%, 275/573) was significantly higher than that in 3-5 years old children (26.6%, 213/801) and that under 3 years old children (18.3%, 270/1480). The difference showed statistical significance (χ(2) = 187.432, P < 0.01). A total of 219 (7.7%) cases were positive for RSV,including 108 RSV-A positive samples and 112 RSV-B positive samples (1 co-infection). The epidemic of RSV showed an obvious seasonal pattern with peaks in autumn,winter and spring,which accounted for 96.8% (212/219) of all the cases; however, the annual incidence of RSV fluctuated greatly. The predominant subtype shifted every 2 years. RSV-B predominated during September 2008 and May 2009, December 2009 and March 2010, accounting for 76.6% (36/47) and 96.9% (62/64) respectively. RSV-A predominated during November 2010 and March 2011, September 2011 and April 2012, accounting for 92.5% (37/40) and 100.0% (48/48) respectively. With the increase of the age, the positive rate of RSV-A and RSV-B decreased gradually (RSV-A: χ(2) = 36.223, P < 0.01; RSV-B: χ(2) = 36.281, P < 0.01). The positive rates of RSV-A in children < 1,1,2,3,4,5-9 and 10-14 years old were 7.0% (26/373), 5.9% (39/662), 4.0% (18/445), 3.2% (13/406), 1.3% (3/236), 1.4% (7/517) and 0.9% (2/215) respectively; while, the positive rates of RSV-B in each age group were 6.4% (24/373), 6.0% (40/662), 4.5% (20/445), 4.4% (18/406), 1.3% (3/236), 1.0% (5/517) and 0.9% (2/215) respectively. The children aged 0-3 years old were more susceptible for RSV infection,accounting for 90.0% (197/219) of the total positive samples. During the outbreak of influenza A H1N1 in November 2009, the positive rate of RSW was 3.0% (3/100), lower than that in the same month of 2008, 2010 and 2011,which were separately 18.2% (6/33), 10.8% (10/93) and 10.0% (4/40). The difference showed statistical significance (χ(2) = 8.450, P < 0.05). During the outbreak of influenza A (H1N1) in January 2011,the positive rate of RSV was 5.7% (3/53), lower than those in the same month of 2009, 2010 and 2012, which was separately 21.7% (5/23), 28.6% (22/77) and 16.0% (8/50). The difference showed statistical significance (χ(2) = 11.233,P < 0.05). During the period of less influenza happened in September 2011, the RSV positive rate was 25.0% (10/40), higher than those in the same month of 2008, 2009 and 2010, which was separately 11.4% (4/35), 1.7% (2/118) and 0.0% (0/109). The difference showed statistical significance (χ(2) = 32.521, P < 0.01).

CONCLUSIONBoth influenza virus and RSV were important etiological agents of ILI of children in Wuhan. The characteristics of seasonal and age distributions of the two viruses were notably different; meanwhile, a certain inhibitional effect of influenza virus on RSV could be observed.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Influenza, Human ; epidemiology ; Male ; Orthomyxoviridae ; classification ; isolation & purification ; Respiratory Syncytial Virus Infections ; epidemiology ; Respiratory Syncytial Viruses ; classification ; isolation & purification

Contraction of smooth muscle cells is triggered by an increase in cytosolic Ca(2+) upon agonist stimulation. Ca(2+) influx across the plasma membrane constitutes a major component of the agonist-induced response in smooth muscle cells. Traditionally, voltage-operated Ca(2+) channel (VOCC) is considered as the channel mediating the Ca(2+) entry. However, this view has been challenged by recent discoveries, which demonstrated that other types of ion channels, such as store-operated and/or receptor-operated Ca(2+) channels (SOCC and/or ROCC), also participate in Ca(2+) response induced by agonists in smooth muscle cells. SOCC is defined as the channel activated in response to the depletion of the internal Ca(2+) stores, an event secondary to G protein coupled receptor or receptor tyrosine kinase stimulation. The Ca(2+) flow mediated by SOCC is termed as capacitative Ca(2+) entry (CCE). Previous study from other group has demonstrated that VOCC played a predominant role in ACh-induced contraction of distal colon smooth muscle in guinea pig. However, whether SOCC participates in the agonist-induced contractile response in this particular tissue is unknown. The present study was performed to investigate the role of CCE in ACh-induced mechanical activity of distal colon smooth muscle in rats. The contractile function of the smooth muscle was assessed by measuring isometric force of isolated rat distal colon rings. We showed that both high extracellular K(+) (40 mmol/L) and ACh (5 mumol/L) evoked striking contraction of the smooth muscle. The contractile responses were almost abolished by removal of extracellular Ca(2+) with ethylene glycol-bis(2-aminoethylether)-N,N,N',N' tetraacetic acid (EGTA), suggesting a critical contribution of extracellular source of Ca(2+) to the contraction. Verapamil (5 mumol/L), an L-type VOCC blocker, significantly attenuated, but didn't completely eliminate the high K(+)- and ACh-induced contraction (74% and 41% for high K(+) and ACh, respectively), indicating that additional channels might be involved in the contractile mechanism. Furthermore, ACh only induced transient contractions in the absence of extracellular Ca(2+). Readmission of Ca(2+) into the extracellular compartment resulted in a significant and sustained increase in the tension of the smooth muscle. This response was not affected by verapamil (5 mumol/L) and Cd(2+) (5 mumol/L), both of which efficiently block VOCC at the doses. However, La(3+), a known inhibitor of SOCC, significantly suppressed the Ca(2+) readdition-induced contraction in a dose-dependent manner. On the basis of these results, we conclude that contraction of smooth muscle in the distal colon is regulated by multiple Ca(2+) channels. In addition to VOCC-mediated Ca(2+) influx, SOCC-mediated CCE participates in agonist-induced contractile response of distal colon smooth muscle in rats.
Acetylcholine ; physiology ; Animals ; Calcium ; metabolism ; Calcium Channels ; physiology ; Colon ; physiology ; Female ; Male ; Muscle Contraction ; physiology ; Muscle, Smooth ; physiology ; Myocytes, Smooth Muscle ; physiology ; Rats ; Rats, Sprague-Dawley ; Verapamil ; pharmacology

OBJECTIVETo determine the jaw bone stress variation affected by cylinder implant diameter and length simultaneously in Ansys DesignXplorer module.

METHODSFinite element model of segment mandible with a cylinder implant was created. The range of the implant diameter (D) and length (L) were set from 2.5 mm to 5.0 mm and from 6.0 mm to 16.0 mm respectively. The maximum Von Mises stresses in jaw bone and sensitivity to D and L were evaluated.

RESULTSUnder axial (buccolingual) load, when one variable equaled to median, the amplification of maximum Von Mises stresses in cortical bone and cancellous bone were 44.66% (71.32%) and 51.45% (58.50%) respectively with the D increasing. The amplification of maximum Von Mises stresses in cortical bone and cancellous bone were 45.97% (21.66%) and 52.15% (37.75%) respectively with the L increasing. When D exceeded 3.7 mm and L exceeded 10.0 mm, the response curve curvatures of maximum Von Mises stresses to L and D in jaw bone ranged from -1 to 0. And the variation of the maximum Von Mises stresses in jaw bone was more sensitive to D than to L.

CONCLUSIONStresses in jaw bone under buccolingual and axial load are apt to be affected by implant diameter and length respectively. And to a cylinder implant, the diameter exceeds 3.7 mm and length exceeds 10.0 mm are optimal selections. Diameter should pay more attention to than to length for cylinder implant. Expanding the width of the jaw bone is more important than expanding the height of the jaw bone in clinical experience.

Biomechanical Phenomena ; Dental Implants ; Dental Prosthesis Design ; Finite Element Analysis ; Humans ; Mandible ; Stress, Mechanical

OBJECTIVETo clone the human tissue factor pathway inhibitor-2 (hTFPI-2) gene and express it by using prokaryotic expression system.

METHODSThe hTFPI-2 coding region was obtained by RT-PCR from human placenta total RNA. The coding fragment was then inserted into prokaryotic expression vector pET19b and expressed in E. coli BL21 by IPTG induction. The produced inclusion bodies were dissolved by denaturalizing chemicals, purified by ion exchange chromatograph, and refolded in air to form proper disulfide bonds. Chromogenic and gelatin zymography methods were used to evaluate the inhibiting effects of hTFPI-2 on trypsin, plasmin and MMPs individually. The inhibitory activity of hTFPI-2 on fabrisarcoma was investigated by matrigel.

RESULTSThe coding fragment of hTFPI-2 was cloned successfully and the protein was expressed as inclusion bodies which account for 20% - 30% of total host protein. The refolded hTFPI-2 could inhibit the invasive ability of fibrisarcoma HT-1080 as well as activity of plasmin, trypsin and MMPs.

CONCLUSIONSThe activated hTFPI-2 is obtained by using prokaryotic expressed system effectively.

Cloning, Molecular ; Escherichia coli ; metabolism ; Gene Expression ; Glycoproteins ; biosynthesis ; genetics ; Humans ; Placenta ; cytology ; RNA ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction

BACKGROUNDIt is well known that conventional splenectomy, which requires careful handling and ligation of tissue of the splenic hilum, can easily cause complications such as splenic fever and pancreatic fistula. Here, we use the technique of dissection of the secondary branches of the splenic pedicle to handle the hilum in the portal hypertension patients who are subjected to splenectomy.

METHODSWe retrospectively compared and analyzed the complications, postoperative hospital stay, operative time, and occurrence of hemorrhage in 121 patients with portal hypertension undergoing splenectomy and devascularization of the gastric cardia from January 1999 to December 2007. The selected cases consisted of 51 patients undergoing conventional splenectomy and 70 patients undergoing dissection of secondary branches of the splenic pedicle. In addition, we analyzed the relationship between size of the spleen and occurrence of complications.

RESULTSThe incidence of pancreatic fistula and splenic fever (0/70 and 9/70) was lower in patients undergoing dissection of secondary branches of the splenic pedicle as compared with that of the conventional group (5/51 and 18/51 respectively). In addition, there was no significant difference in operative time and volume of blood loss between two groups. The spleen thickness of those patients who had pancreatic fistula and splenic fever was significantly greater than those without complications.

CONCLUSIONSThese results indicate that dissection of secondary branches of the splenic pedicle in portal hypertension patients undergoing splenectomy can decrease the incidence of splenic fever and pancreatic fistula, and shorten the postoperative hospital stay, especially in the patients with a large spleen. So dissection of secondary branches of the splenic pedicle is a valuable technique for splenectomy.

Adolescent ; Adult ; Aged ; Female ; Humans ; Hypertension, Portal ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Spleen ; blood supply ; surgery ; Splenectomy ; methods ; Treatment Outcome ; Young Adult