Volume 124 ·  Issue 21 ·  2011

Previously,hydrogen sulfide (H2S) was considered to be a toxic gas.However,recently it was discovered that it could be produced in mammals and even in plants,throughtheproductionandmetabolismof sulfur-containing amino acids.In mammals,H2S is mainly catalyzed by cystathionine-γ-lyase (CSE),cystathionin-β-lyase (CBS) and 3-mercaptopyruvate sulfurtransferase (MPST) with the substrate of L-cysteine.Endogenous H2S exerts many important physiological and pathophysiological functions,including hypotensive action,vasorelaxation,myocardial dilation,inhibition of smooth muscle cell proliferation,and antioxidatve actions.Importantly,it plays a very important role in the pathogenesis of systemic hypertension,pulmonary hypertension,atherosclerosis,myocardialinjury,angiogenesis,hyperhomocysteinemi aandshock.Therefore,H2S is now being considered to be a novel gasotransmitter after nitric oxide and carbon monoxide in the regulation of circulatory system。

Abstract：Background Cardiac involvement is the most common complication of Kawasaki disease (KD); however,the underlying mechanisms are not understood.The present study was designed to investigate changes in plasma hydrogen sulfide (H2S) and nitric oxide (NO) levels in the acute and recovery stages of KD children and to examine their clinical significance.Methods Thirty-five KD patients and 32 healthy children were enrolled in the study.KD patients were divided into two subgroups:a non-cardiac involvement group and a cardiac involvement group.Plasma H2S levels were measured using the sulfur-sensitive electrode method and plasma NO levels and NO synthase activity were determined using the nitrate reductase method both before and after intravenous immune globulin (IVIG) therapy.Results Plasma H2S levels significantly decreased in KD patients during the acute phase of the disease and NO levels were significantly increased,compared with the control group (P ＜0.01).After treatment with IVIG,both plasma H2S and NO levels significantly increased (P ＜0.01).The plasma levels of H2S were significantly lower in the cardiac involvement group compared with the non-cardiac involvement group (P＜0.05).Conclusion H2S and NO may play a role in the pathophysiological process of inflammation during the acute phase of KD.Endogenous H2S may exert protective effects with respect to cardiac complications in KD。

Abstract：Background Central nervous system leukemia (CNSL) is an important relapse in children with acute lymphoblastic leukemia (ALL).We investigated the possible role of endogenous hydrogen sulfide (H2S) of cerebrospinal fluid (CSF) in predicting CNSL.Methods From August 2008 to December 2010,380 children were enrolled in this study at Shijitan Hospital,China.These children were from 2 to 16 years old,and the median age was 6.5 years.They were divided into a CNSL group (7 cases),a leukemia group (307 cases),a non-leukemia group (26 cases) and a healthy group (40 children).CSF specimens were obtained from conventional lumbar punctured,then centrifuged and supernatants preserved for H2Sdetection.Leukemic cells precipitates from CSF were found in three cases,the hCSE and hCBS mRNA expression was detected by reverse transcription polymerase chain reaction (RT-PCR),and H2S levels in serum were also measured.The receiver operating characteristic (ROC) curve and area under curve (AUC) were used to assess the predictive diagnosis role of CSF H2S in children with ALL and CNSL.Results The serum H2S contents of the CNSL and leukemia groups were (96.98+15.77) μmol/L and (93.35+17.16)μmol/L respectively,much higher than those of healthy,(44.29+2.15) μmol/L,and non-leukemia,(46.32±6.54) μmol/L,groups (P ＜0.01).Compared with the leukemia group,CSF H2S content of the CNSL group was significantly high (P ＜0.01).Meanwhile,in contrast to the non-leukemia group,CSF H2S contents of the CNSL and leukemia groups were both significantly increased (P ＜0.01).In addition,leukemic cells from CSF precipitations could express CBS and CSE mRNA.Furthermore,the ROC analysis showed the UAC was 0.929 (95％ Cl:0.857-1.000),and the optimum cut-off value of CSF H2S was 12.08μmol/L,and the sensitivity and specificity were 83.3％ and 97.2％ respectively.Conclusions CSF H2S contents were significantly increased in children with CNSL.After treatment,H2S contents were decreased subsequently.Therefore,we speculated that H2S levels of CSF would predict CNSL in ALL children。

Abstract：Background It has been reported that endogenous or exogenous hydrogen sulfide (H2S) exerts physiological effects in the vertebrate cardiovascular system.We have also demonstrated that H2S acts as an important regulator of electrophysiological properties in guinea pig papillary muscles and on pacemaker cells in sinoatrial nodes of rabbits.This study was to observe the electrophysiological effects of H2S on human atrial fibers.Methods Human atrial samples were collected during cardiac surgery.Parameters of action potential in human atrial specialized fibers were recorded using a standard intracellular microelectrode technique.Results NaHS (H2S donor) (50,100 and 200 μmol/L) decreased the amplitude of action potential (APA),maximal rate of depolarization (Vmax),velocity of diastolic (phase 4) depolarization (VDD) and rate of pacemaker firing (RPF),and shortened the duration of 90％ repolarization (APD90) in a concentration-dependent manner.ATP-sensitive K+ (KATP)channel blocker glibenclamide (Gli,20 μmol/L) partially blocked the effects of NaHS (100 μmol/L) on human atrial fiber cells.The L-type Ca2+ channel agonist Bay K8644 (0.5 μmol/L) also partially blocked the effects of NaHS (100 μmol/L).An inhibitor of cystathionine y-lyase (CSE),DL-propargylglycine (PPG,200 μmol/L),increased APA,Vmax,VDD and RPF,and prolonged APD90.Conclusions H2S exerts a negative chronotropic action and accelerates the repolarization of human atrial specialized fibers,possibly as a result of increases in potassium efflux through the opening of KATP channels and a concomitant decrease in calcium influx.Endogenous H2S may be generated by CSE and act as an important regulator of electrophysiological properties in human atrial fibers。

Abstract：Background Atherosclerosis is an important cardiovascular disease,becoming a major and increasing health problem in developed countries.However,the possible underlying mechanisms were not completely clear.In 2009,our research group first discovered that hydrogen sulfide (H2S) as a novel gastrotransmitter played an important anti-atherosclerotic role.The study was designed to examine the regulatory effect of hydrogen sulfide (H2S) on endoplasmic reticulum stress (ERS) in apolipoprotein E knockout (apoE(-/-)) mice fed a Western type diet.Methods C57BL/6 mice and homozygous apoE(-/-) mice were fed a Western type diet.C57BL/6 mice were injected intraperitoneally with normal saline (5 ml/kg per day) as control group.The apoE+ mice were treated with the same dose of normal saline as the apoE(-/-) group,injected intraperitoneally with sodium hydrosulfide (NaHS,an H2S donor,56μmol/kg per day) as the apoE(-/-)+NaHS group and injected intraperitoneally with DL-propargylglycine (PPG,a cystathionine-y-lyase inhibitor,50 mg/kg,per day) as the apoE/ +PPG group.After 10 weeks,the mice were sacrificed and the plasma lipids were detected.Sections of aortic root from these animals were examined for atherosclerotic lesions by HE and oil red O staining.The aortic ultrastructure and microstructure were analyzed with the help of light and electronic microscope.Glucose-regulated protein 78 (GRP78),caspase-12,copper-andzinc-containing superoxide dismutase (Cu/ZnSOD) and Mn-containing superoxide dismutase (MnSOD) protein expression in aortic tissues were detected with immunohistochemistry.The level of intracellular reactive oxygen species (ROS) were measured by using a commercial assay kit.Results Compared with control mice,apoE(-/-) mice showed increased plasma levels of total cholesterol (TC),triglyceride (TG) and low density lipoprotein (LDL),decreased high density lipoprotein (HDL),increased aortic plaque size,destroyed ultra-structure of aortic tissue,and increased expression of GRP78 and caspase-12 proteins.Compared with apoE(-/-) mice,H2S donor-treated apoE(-/-) mice showed a decreased plasma LDL level,lessened plaque necrosis and attenuated aortic ultra-structural disorder.H2S donor-treatment induced GRP78 expression but suppressed caspase-12 expression in aortic lesions.However,compared with apoE/ mice,PPG treated apoE(-/-) mice showed enlarged plaque size,more severe ultrastructural disorder of the aortic tissue and reduced GRP78 staining in aortic lesions.The plasma lipids and the staining of caspase-12 in apoE/ + PPG rats did not significantly differ from those in the apoE(-/-)mice.Consistently,H2S induced SOD expression,accompanied by a reduced level of ROS.Conclusion H2S plays a regulatory role in aortic ERS and reduces atherosclerotic lesions in apoE(-/-) mice fed with a Western type diet。

Abstract：Background Endogenous hydrogen sulfide is a new neuromodulator which takes part in the regulation of central nervous system physiology and diseases.Whether endogenous hydrogen sulfide in the central nervous system regulates cardiovascular activity is not known.In the present study,we observed the hemodynamic changes of hydrogen sulfide or its precursor by intracerebroventricular injection,and investigate the possible roles of endogenous digitalis like factors and sympathetic activity in the regulation.Methods Ninety-four Sprague-Dawley rats underwent a right cerebroventricular puncture,then the hydrogen sulfide saturation buffer or its precursor injected by intrcerebroventricular catheter.A heperin-filled catheter was inserted into the right femoral artery or into the left ventricle,and changes of blood pressure or cardiac function recorded by a Powerlab/4S instrument.Phentolamine or metoprolol were pre-injected to observe the possible role in autonomic nerve activity.After rats were sacrificed,plasma was collected and endogenous digitalis-like factors were measured with a commercial radioimmunoassay kit.The aortic,cardiac sarcolemmal vesicles were isolated and the activity of Na+-K+-ATPase was measured as ouabain-sensitive ATP hydrolysis under maximal velocity conditions by measuring the release of inorganic phosphate from ATP.Unpaired Student's ttest for two groups or analysis of variances (ANOVA) for multiple groups were used to compare the differences of the changes.Results Intracerebroventricular injection of hydrogen sulfide induced a transient hypotension,then dramatic hypertenive effects in a dose-dependent manner.Bolus injection of L-cysteine or beta-mercaptopyruvate also increased mean arterial pressure (P ＜0.01),whereas hydroxylamine-a cystathionine beta synthase inhibitor decreased the arterial pressure (P ＜0.01).Hydrogen sulfide and L-cysteine increased mean arterial pressure,left ventricular develop pressure and left-ventricle maximal rate of systolic and diastolic pressure; these functions were decreased by hydroxylamine (P ＜0.01).Glibenclamide (a KATP channel blocker) blocked the transient hypotensive effect,phentolamine (an alpha-adrenergic receptor blocker) blocked the hypertensive effect,and metoprolol (a selective beta 1 receptor blocker) blocked the positive inoptropic effect of central nervous system hydrogen sulfide.The endogenous digitalis-like factors in plasma were elevated (P＜0.01) after treatment with L-cysteine,association with decreasing Na+-K+-ATPase activity in cardiac or aortic sarcolemmal vesicles (P ＜0.01).Hydroxylamine injection reduced the endogenous digitalis-like factors level in plasma association with increasing Na+-K+-ATPase activity in cardiac and aortic sarcolemmal vesicles.Conclusion Central nervous system endogenous hydrogen sulfide upregulated mean arterial pressure and cardiac systolic function by activation of sympathetic nerves or release of endogenous digitalis-like factors。

Abstract：Background Acute lung injury (ALl) is a common critical disease in emergency care.Oxidative stress plays an important role in the pathogenesis of ALl.Endogenous hydrogen sulfide (H2S) can inhibit oxidative stress in rat gastric mucosal epithelium.In this study,we examined the possible role of H2S in regulation of the oxidative stress in oleic acid-induced acute lung injury in rats.Methods The rat model of ALl was induced by intra-tail vein injection of oleic acid (OA).NaHs solution was injected intraperitoneally before OA injection as an OA+NaHS group.A semi-quantitative histological index of quantitative assessment of lung injury was calculated.The levels of superoxide dismutase (SOD),glutathione (GSH) and malondialdehyde (MDA) in plasma and lung tissue were detected with ELISA.The levels of H2S content in lung tissue were detected with an ion meter.Results Compared with the control group,the level of H2S in lung tissue was significantly decreased,and the level of SOD and GSH were decreased but the level of MDA was increased in plasma and lung tissue in rats with ALl.NaHS lessened the ALl in association with an increase in the level of H2S in lung tissue,a decrease in the level of MDA but an increase in SOD and GSH levels in plasma and lung tissues.Conclusion Endogenous H2S could inhibit the oxidative stress in lung tissue in oleic acid-induced acute lung injury inrats。

Abstract：Background Classically,myomectomy has been performed via laparotomy,but laparoscopic myomectomy has now become a valuable treatment option.Vaginal myomectomy as a minimal invasive procedure has also been evaluated; however,its feasibility and safety are controversial with few clinical trials published.This study was designed to evaluate the feasibility of vaginal myomectomy in China and to document any associated complications.Methods From January 2005 to December 2010,43 patients with symptomatic myomas were admitted for vaginal myomectomy in Peking Union Medical College Hospital (PUMCH).The indications,operative performance,postoperative complications and outcome of these patients were analyzed retrospectively.Results Myomectomy was performed vaginally in all patients.The mean operating time was (66.4 + 22.6) minutes and the mean operative blood loss and hospital stay were (78.3±64.4) ml and (4.9±3.3) days,respectively.Five (12％) patients developed febrile morbidity and experienced a high postoperative temperature (mean 38.4℃).Postoperative fever was associated with greater operative trauma,longer operative time and greater weight of the tumor (all P ＜0.01).The cost of surgery was RMB (820.6+339.1) Yuan ((124.3±51.4) US dollars) and the total medical cost was RMB (4880.4±1088.4) Yuan ((739.5±164.9) US dollars).Three patients later conceived spontaneously and had uneventful vaginal deliveries.Only one patient had a tumor recurrence during the following-up period.Conclusion Vaoinal mvomectomv is a feasible and safe surgical procedure with low recurrence and complication rates。

Abstract：Background Food composition and style is changing dramatically now,which causes inappropriate secretion of hormones from brain,gastrointestinal and endo-pancreas,may be related to unbalance of glucose in blood.The aim of this study was to explore the fast response of C-peptide,glucagon-like peptide-1 (GLP-1),ghrelin and endomorphin-1 (EM-1) to the eastern and western style meals in patients with type 2 diabetes mellitus.Methods The study enrolled 57 patients with type 2 diabetes (20 men and 37 women,mean age (67.05±8.26) years).Eastern style meal (meal A) and western style meal (meal B) were designed to produce the fullness effect.C-peptide,GLP-1,ghrelin and EM-1 were assessed before (0 hour) and after (2 hours) each diet.Results The delta (2h-0h) of C- peptide in meal A was significantly lower than that in meal B (P=0.0004).C-peptide,GLP-1,ghrelin and EM-1 were obviously higher before meal B than those before meal A (P ＜0.0001,＜0.0001,=0.001,=0.0004 respectively).Blood glucose 2 hours and 3 hours after meal B were higher than those after meal A (P=-0.0005,0.0079 respectively).Correlations between GLP-1 and ghrelin were strongly positive before both meals and 2 hours after both meals and also in relation to the delta of meal A and meal B (rA0h=0.7836,rB0h=0.9368,rAsh=0.7615,rB2h=0.9409,rA(2h_0h)=0.7531,rB(2h-0h)=0.9980,respectively,P ＜0.0001).Conclusion Western style meal (high fat and protein food) could make more response of C-peptide than eastern style meal,and could stimulate more gut hormones (GLP-1,ghrelin) and brain peptide (EM-1) at the first phase of digestion。

Abstract：Background Streptococcus (S.) oligofermentans is a newly identified bacteria with a yet to be defined mechanism of sucrose metabolism that results in acid production.This study aimed to investigate the biochemical mechanisms of S.oligoferm-entans glucose metaolism.Methods The S.oligofermentans LMG21532,Lactobacillus (L.) fermentum 38 and the S.S.mutans UA140 were used to characterize sucrose metabolism by measuring lactate dehydrogenase (LDH) activity and lactic acid production.Continuous dynamics and high performance capillary electrophoresis were used to determine LDH activity and lactic acid production,respectively,from bacteria collected at 0,10 and 30 minutes after cultured in 10％ sucrose.Results These analyses demonstrated that LDH activity of the three bacterial strains examined remained stable but significantly different throughout the sucrose fermentation process.The S.o/igofermentans LDH activity ((0.61±0.05) U/mg) was significantly lower than that of L.fermentum ((52.91+8.97) U/mg).In addition,the S.oligofermentans total lactate production ((0.048±0.021) mmol/L) was also significantly lower than that of L.fermentum ((0.958±0.201) mmol/L).Although the S.oligofermentans LDH production was almost double of that produced by S.mutans ((0.32±0.07) U/mg),lactic acid production was approximately one sixth that of S.mutans ((0.296±0.058) mmol/L).Additional tests examining pyruvic acid production (the LDH substrate) demonstrated that lactic acid concentrations correlated with pyruvic acid production.That is,pyruvic acid production by S.oligofermentans was undetectable following sucrose incubation,however,(0.074±t0.024) and (0.175±0.098) mmol/L pyruvic acid were produced by S.mutans and L.fermentum,respectively.Conclusion S.oligofermentans is incapable of fermenting carbohydrates to produce enough pyruvic acid,which results in reduced lactic acid production。

Abstract：Background Cyclosporine is effective in treating nephrotic syndrome (NS) with idiopathic membranous nephropathy (IMN) in adults.But high relapse rate remains a major concern.The way to manipulate cyclosporine is inconclusive.The aim of this study was to introduce the way how to titrate the cyclosporine to maintain complete remission without relapse.Methods Patients with biopsy-proven IMN with NS treated with cyclosporine for at least 1 month from 1996 to 2011 at Peking Union Medical College Hospital were reviewed.Results Mean age of the 51 eligible patients was 52 years,with 39 men.Mean proteinuria was (7.47+3.14) g/d,serum albumin (24.50±6.29) g/L,and serum creatinine (82.62+21.18) μmol/L.Cyclosporine was commenced at a mean dose of (3.46±0.63) mg.kg1.d-1.Ora1 prednisone (0.40±0.29) mg·kg1·d-1 was given concomitantly in 38 patients.Cyclosporine was administered for a median of 16 months (range 1-93 months) and stopped in non-responders by month six.By month 3 (n=47),the number in complete remission (CR) and partial remission (PR) was 3 and 24,which shifted to 12 and 17 by month 6 (n=41).Male gender,heavy proteinuria,low serum albumin level,and high serum creatinine level were significant determinants in poor response by month six (P ＜0.05 in all variables compared with responders).There was a significant reversible serum creatinine increase within 25％ during month 3 to 12 (P ＜0.05 in all variables compared with baseline value).Eleven patients maintained cyclosporine for more than 24 months with a cyclosporine dose of (1.04±1.06)mg·kg1·d-1.Nine patients were in CR.Renal function,systolic and diastolic blood pressure remained stable.Renal impairment (＞30％ rise of serum creatinine),secondary infection,hypertension,gingival hyperplasia and liver impairment occurred in 6,4,10,4,and 1 patients,respectively.Conclusions The observation time for cyclosporine to effectively induce CR of NS in IMN adults should be at least six months.Long-term and low-dose of cyclosporine therapy is safe and effective to maintain CR in those responders。

Abstract：Background Video-assisted thoracic sympathetic block is an effective,safe,and minimally invasive method fortreatment of primary hyperhidrosis.The purpose of this study was to decide which one of using electrocautery hook and titanium clip is the appropriate procedure for primary palmar hyperhidrosis by assessing the compensatory sweating (CS)and quality of life (QOL) of patients after sympathetic block.Methods Between October 2007 to August 2010,120 patients with primary palmar hyperhidrosis were randomly divided into two groups,electrocautery hook group (60 patients) and titanium clip group (60 patients).All patients were treated by sympathetic block at T4 level.The CS was graded based on severity and location; the QOL was classified to 5 different levels based upon the summed total scores (range from 20 to 100) before and after surgery.The variables were compared.Results The postoperative follow-up period was 2 months.All patients were cured.Three patients in electrocautery hook group and 1 patient in titanium clip group had a unilateral pneumothorax on chest X-ray,but none of them was necessary to have chest drainage.Neither perioperative mortality nor serious complications such as cardiac arrhythmia or arrest were observed during the operation.No bradycardia or Horner's syndrome occured.CS was not more common in patients in titanium clip group than in those in electrocautery hook group (P=0.001).Moderate and severe CS was few in all patients,and there was no significant difference between two groups (P=-0.193).Most of the patients feel a notable improvement of the the QOL; nevertheless,there was no significant difference between the groups (P=0.588).Conclusions Both electrocautery hook and titanium clip used for sympathetic block at the T4 level are effective,safe,and minimally invasive for palmar hyperhidrosis.Because of the lower severity of CS and the similar improvements in the QOL after operation,we prefer to use of titanium clip for treating palmar hyperhidrosis。

Abstract：Background Several studies have suggested that the exposure to cadmium (Cd) increased mortalities from renal diseases,cardiovascular diseases and malignant neoplasm,including lung cancer and prostate cancer among inhabitants living in Cd-polluted areas and factory workers.This study aimed to assess the influence of environmental exposure to Cd on long term outcome of inhabitants living in an area polluted by Cd.Methods A 22-year follow-up study was conducted with 3119 inhabitants (1403 men and 1716 women) living in the Cd polluted Kakehashi River basin in Japan.The subjects were divided into 4 groups according to the amount of urinary Cd level (＜3.0 μg/g creatinine (Cr),3.0-4.9 μg/g Cr,5.0-9.9 μg/g Cr,and ≥10.0 μg/g Cr).Mortality was calculated by the person-years method.Hazards ratios (HR) and 95％ confidence intervals (C/) were assessed by the Cox's proportional hazard model.Results Compared with urinary Cd ＜3.0 μg/g Cr group,the HR of 5.0-9.9 μg/g Cr and ＞10.0 μg/g Cr groups were significantly increased after adjustment for age in both sexes:1.24 (95％ CI 1.01-1.51) and 1.48 (95％ CI 1.17-1.90) for men; 1.64 (95％ C/ 1.17-2.28) and 1.78 (95％ C/ 1.27-2.50) for women.The most frequent cause of death was malignant neoplasm in men and cardiovascular diseases in women.The significant increase in mortality risk for cardiovascular diseases was observed in the subjects with ≥10 μg/g Cr in both sexes:1.79 for men (95％ Cl 1.02-3.12) and 2.38 for women (95％ CI 1.11-5.07).When the subjects were divided into 2 categories (＜20 μg/g Cr and ＞20 μg/g Cr),the HR of the urinary Cd ≥20 μg/g Cr group for nephritis and nephrosis were 4.82 (95％ CI 1.07-21.61) in men and 7.92 (95％ Cl 1.77-35.33) in women,respectively.The significant increase was not observed for malignant neoplasm.Conclusion These results suggest a dose-response relationship between Cd body burden and mortality for cardiovascular diseases,cerebrovascular diseases and nephritis and nephrosis。

Abstract：BackgroundPolymer coating on coronary stents induces vascular inflammatory response,reduces re-endothelialization,and affects long-term outcome after percutaneous coronary intervention (PCI).The SERY-1 registry aimed to determine whether a novel polymer-free paclitaxel-eluting microporous Yinyi stent could improve 1-year outcome after index procedure in real-world clinical practice.Methods Clinical and angiographic data and follow-up outcome were collected in 1045 patients who underwent PCI with implantation of ≥1 Yinyi stents between June 2008 and August 2009 at 27 medical centers.The primary endpoint was the cumulative rate of composite major adverse cardiac events (MACE) and the secondary endpoint was the incidence of stent thrombosis at 1 year.Results Overall,1376 lesions were treated successfully with 1713 Yinyi stents,and 1019 (98.7％) patients received dual antiplatelet therapy for at least 12 months.During 1-year follow-up,8 patients (0.78％) had cardiac death,6 (0.58％)suffered non-fatal myocardial infarction,and 46 (4.46％) underwent repeat PCI due to recurrence of angina,resulting in 1-year MACE-free survival of 94.09％.Stent thrombosis occurred in 10 (0.97％) patients,and the rate of Academic Research Consortium (ARC) definite or probable stent thrombosis was 0.78％.Conclusions Polymer-free paclitaxel-eluting microporous Yinyi stent is effective and safe for interventional treatment of coronary artery disease in real-world clinical practice,without recourse to carrier polymer.Potential long-term clinical advantages of this stent deserve further investigation。

Abstract：Background Murine cytomegalovirus (MCMV) early protein M112-113 is involved in viral DNA replication and believed to play a crucial role in the viral pathogenesis.To investigate the biological function of M112-113 protein in the pathogenesis of the brain disorders caused by cytomegalovirus (CMV),a screening for proteins interacting with M112-113 was performed by a yeast two-hybrid system.Methods Bait plasmid pGBKT7-M112-113 was constructed and transformed into AH109 yeast.After confirmation of the expression of MCMV M112-113 in yeast,the bait yeast was mated with a prey yeast containing mouse brain cDNA library plasmid to screen the proteins interacting with M112-113.Interactions between M112-113 and the obtained proteins were verified by yeast two-hybrid assay and chemiluminescent co-immunoprecipitaion.Results Two proteins interacting with M112-113 were identified,including metastasis-associated 1 (MTA1) and zinc finger,CCHC domain containing 18 (ZCCHC18).M112-113 protein could interact with MTA1 or ZCCHC18 in yeast and mammalian cells.Conclusion The interactions of M112-113 with MTA1 or ZCCHC18 may be related to the pathogenesis of MCMV-associated disease in central nervous system。

Abstract：Background Transplantation of mensenchymal stem cells (MSCs) has been proposed as a promising way for tissue engineering.However,the application of MSCs for transplantation will undergo apoptosis due to the extremely harsh microenvironment such as excessive inflammation.Apigenin (API) has been reported to protect cells against inflammatory damage and cell death by exhibiting anti-inflammatory and anti-oxidative capacity.Here we investigated the modulatory effects of API in lipopolysaccharide (LPS)-mediated inflammation and apoptosis of MSCs,and further defined the underlying mechanism.Methods Effects of different concentrations of API (0,5,10,20,40 and 80 μmol/L) for 24 hours,and LPS (0,0.5 and 5.0 μg/ml) for 6 hours and 24 hours on MSCs viability were assayed by MTT.Based on this,MSCs were pretreated with different concentrations of API (0-40 μmol/L) at the indicated times (6,12 and 24 hours) followed by exposure to 5 μg/ml LPS for 24 hours.MTT,phase-contrast microscopy,annexinV/propidium iodide (PI) double stain flow cytometry (FCM) and Hoechst staining were applied to explore the effects of API on MSCs induced by 5 μg/ml LPS for 24 hours.In addition,reverse-transcription polymerase chain reaction (RT-PCR) was applied to detect the mRNA expression of pro-inflammatory factors including cyclooxygenase-2 (COX-2),inducible nitric oxide synthase (iNOS),nuclear factor-kappa B (NF-KB),pro-apoptotic gene caspase-3,Bad,and anti-apoptotic gene Bcl-2.Moreover,AutoDock software was used to imitate the docking score of API and vitamin D receptor (VDR).In parallel,Western blotting and RT-PCR were used to investigate protein and mRNA expression of VDR.Results MSCs stimulated with LPS 5 μg/ml for 24 hours was used as a model of apoptosis induced by over inflammatory stimulus.API (0-40 μmol/L) had non-toxic effect on MSCs; however,it could decrease mRNA expression of COX-2,iNOS and NF-KB at different time points in MSCs induced by LPS,except for API at the concentration of 5 μmol/L.Results from phase-contrast microscopy,MTT,Hoechst staining and AnnexinV/PI double stain FCM demonstrated that with the increasing concentrations of API and extension of administrating time,significant morphological changes of MSCs occurred,viability of cells was strongly inhibited,and meanwhile,apoptosis of LPS-administrated MSCs was exacerbated,compared with LPS individual group.In addition,API promoted caspase-3,Bad mRNA expression and inhibited Bcl-2 mRNA expression in a time-dependent and concentration- dependent manner.Further study found that pro-apoptosis effect of API was related to suppress VDR expression.Conclusions API could inhibit the expression of inducible inflammatory factors,therefore exert the strong anti-inflammatory function.However,API could not protect MSC apoptosis induced by LPS but amplified the apoptosis.The apoptosis is related to Bad/Bcl-2 increasing and caspase-3 activation,which is mediated through suppressing VDR expression。

Abstract：Background The prevalence of chronic pancreatitis has increased during recent years in Asia-Pacific areas as well as in China.The etiologies vary in different regions and periods.This study aimed to investigate the changing etiologies of chronic pancreatitis within 20 years at Peking Union Medical College Hospital in China.Methods Retrospective analysis of the etiologies of 636 cases of chronic pancreatitis at Peking Union Medical College Hospital from 1990 to 2010 was performed.Patients were divided into two groups according to two time periods (1990-2000 and 2001-2010).Statistical analysis was performed using the chi-square test.Results The morbidity rate of chronic pancreatitis in China has recently increased.The main etiology changed from biliary diseases in the 1990s (decreased from 36.8％ to 28.1％) to alcohol abuse after the year 2000 (increased from 26.5％ to 36.8％).The main etiology of biliary diseases is stones in the cholecyst or bile duct,and the percentage of cholecystitis cases has increased.Autoimmune disease,including autoimmune pancreatitis,has increased quickly and currently accounts for 7.3％ of cases because a greater number of autoimmune pancreatitis cases are being diagnosed.Approximately 9.5％ of chronic pancreatitis cases are caused by multiple factors such as alcohol abuse and bile duct stones.Other factors include cholecystectomy and acute pancreatitis.Conclusions The main etiology of chronic pancreatitis has changed from biliary disease to alcohol abuse in recent years.Autoimmune factors have also obviously increased。

Abstract：Background Although CD4+ T cell apoptosis and CD8+ T cell responses have been extensively studied during HIV infection,how apoptosis signals being initiated in CD4+ T cells still need to be elucidated.The present study was designed to characterize the function-unknown gene,C6orf120,and elucidates its primary role in tunicamycin-induced CD4+ T apoptosis.Methods The C6orf120 coding sequence was amplified from peripheral blood mononuclear cells (PBMCs) total RNA of AIDS patients.The DNA fragment was inserted into the pET-32a expression system,transformed into Escherichia coli,and preparation of C6ORF120 recombinant protein.The magnetic cell separation technology was used to prepare primary CD4+ T cells and CD8+ T cells.The primary T cells were cultu red at 1 x 106 cells/ml,treated with 0,0.1,1,10,100,and 200 ng/ml of C6orf120 recombinant protein for 48 hours,then harvested for cell cycle and apoptosis analysis.Tunicamycin (0.5 μmol/L) was used to induce endoplasmic reticulum stress in Jurkat cells.The biomarker 78 KDa glucose-regulated protein (GRp78) and growth arrest and DNA damage (GADD) were used to evaluate endoplasmic reticulum stress of Jurkat cells.Results We prepared C6ORF120 recombinant protein and its polyclonal antibody.Immunohistochemical analysis showed that C6orf120 mainly expressed in hepatocytes and cells in germinal center of lymph node.At concentration of 0.1,1,10,100,and 200 ng/ml,C6orf120 recombinant protein could induce apoptosis of Jurkat cells and primary CD4+T cells,and promoting G2 phase of its cell cycle.Western blotting analysis showed that C6ORF120 recombinant protein increased the expression of GRp78 and GADD in Jurkat cells in vitro.Conclusion Our results suggested that C6ORF120 could induce apoptosis of CD4+ T cells,at least in part,mediated with endoplasmic reticulum stress。

Abstract：Apoptosis is a physiologically essential mechanism of cell and plays an important role in reducing the development and progression of tumors.The appealing strategy for cancer therapy is to target the lesions that induce apoptosis in cancer colls.Survivin,the smallest member of the mammalian inhibitors of the apoptosis protein family,is upregulated in various malignancies to protect cells from apoptosis.Survivin knockdown could induco cancer cell apoptosis and inhibit tumor-angiogenesis.Survivin expression would be silenced by microRNA (miRNA)-mediated RNA interference.However,noninvasive and tissue-specific gene delivery techniques remain absent recently and the utilizations of miRNA expression vectors have been limited by inefficient delivery technique,especially in vivo.On the other hand,safe and promising technologies of gene transfection would be valuable in clinical gene therapy.Successful treatment of gene transfer method would lead to a new and readily available approach in the anticancer research.Sonoporation is an alternative technique of gene delivery that uses ultrasound targeted microbubble destruction to create pores in the cell membrane.Based on our previous studies,in this article,we postulated that the transfection of miRNA could be mediated by the combination of sonoporation and polyethylenimine (PEI) which was one of the most effective poly-cationic gene vectors and enhance the endocytosis of plasmids DNA and hypothesized that the gene silencing and apoptosis induction with miRNA targeting human Survivin would be improved by this novel technique.In our opinion,this novel combination of sonoporation and PEI could enhance targeted gene delivery effectively and might be a feasible,novel candidate for gene therapy。

To the Editor:The National Health Service (NHS) in England launched a national program called National Program for IT (NpflT) in June 2000 after recognizing the need for a comprehensive update of NHS IT systems.The central element of the program is the national electronic health record from 'the cradle to the grave' for every citizen of England.The program was original scheduled (now changed through change and contract amendment) to be rolled out in four phases over a period of eight years,having started in April 2002,and due to end in December 2010。

To the Editor:The apraxia of eyelid opening (ALO) refers to a nonparalytic movement disorder of eyelid characterized bv transient difficulty in voluntarily initiating the act of lid elevation in spite of preserved alertness and language comprehension.1 Thetypical freezing of gait (FOG) is characterized by an abrupt difficulty in starting or continuing rhythmic and repetitive movements.The frequency of ALO was 0.3％-0.7％ in Parkinson's disease (PD),2,3 and it almost concurrently occurred with blepharospasm (BPS) or other dystonia.Yoon et al3 reported 3 ALO cases with BPS out of 913 PD patients.It is rare that a PD patient presents isolated ALO (not with BPS or other dystonia) as well as FOG.We reported here a case of PD with isolated ALO and FOG。