Volume 89  ·   Issue 16  ·  2009

Keloid is the result of abnormal skin wound repair. Although its pathogenesis is still not fully elucidated, it is known that it is closely related to the following factors: (1) abnormal function of fibroblasts, immune cells and keratinocytes; (2) Extracellular matrix abnormalities such as hyaluronic acid, collagen and tendon antigen protein; (3) Abnormal expression of growth factors such as transforming growth factor β (TGF-β) and basic fibroblast growth factor (bFGF); (4) abnormal apoptosis; (5) immunological abnormalities; (6) endocrine abnormalities such as sex hormones; (7) inheritance; (8) mechanical or mechanical factors; (9) Treatment factors such as triamcinolone acetonide, tretinoin and radiation.

Skin scars include physiological scars and pathological scars, and the latter includes hypertrophic scars and keloids. Scar-related diseases not only cause physical diseases, but also directly lead to psychological obstacles in patients. Although the research on scar has been continued for decades, there is still a certain distance to overcome this difficult problem [1]. In recent years, the progress of basic research such as regenerative biology and regenerative medicine has provided some new ideas for in-depth research on scar.

Objective To investigate whether intralesional injection of low-dose 5-fluomuracil(5-Fu)and corticostemids Can increase the effective rate and decrease the recurrence rate of flurgically excised auricular keloid Methotis Eighty-three patients with 166 total ear keloids were studied,including 79 keloids of helix and 87 of earlobe with an average disease history of 2.9 years.Ear piercing accounted for 92.7%for etiology.Keloids were first surgically rcmoved in order to keep the normal auricular shape.Then intrahsional injeefion of a low-dose of 5-Fu and corticosteroids was given 3-4 weeks after surgery and every 4 weeks afterwards.If no recurrence,the injection was given every 2 to 3 months with gradually tapered dose.The evaluation smndard includes:patients with no recurrence 6 months after stopping drug injection and maintaining of the noFmal auricular shape are considered as"cure":those remain having minor auricular deformity 6 months post.operation with no recurrence and needing further treatment are considered as "effective".Results The therapeutic period for 83 patients Was 2-26 months(meas:7 months).The patient follow-ups were 0-49 months(mean:9 monks)after withdrawing drug injection.There were 39 cured cases(47.0%)and 44 effective cases(53%).The total effective rate was 100%.Normal auricular shapes were achieved in 77 cases(92.7%).Conclusion Surgical removal of auricular keloid followed by intralesional injection of low-dose 5-Fu and corticostemid8 is an effective method to treat auricular keloid and prevent its relapse.

Objective To investigate the proper ways to manage a large cícatíical scalp alopeeia.Methodls Based on location,size and condition of scarring Iesiorm,218 patients with 20%-75%scarring alopecia were selected and treated with either dense-packing hair grafting technique(136 cases)or tissue expansion(82 cases).There were 209 in burn,5 in avulsed iniury and 4 in tumor excision.The lesions with tIIick.stable scars in the frontal or/and temporal areas underwent either hair grafting or tissue expansion process as desired by the patients.In addition,the alopecia in an unstable scar on the skull,with skull defect under the lesion and the lesion involved in the back of head was only treated with tissue expansion.The hair-grafting technique was carried out by harvesting a scalp strip from the back of head,then dividing it into a series of 1-3 hair grafts and finally implanting them into the prepared recipient holes of lesion with a desired hair direction.Ten areas with 1 cm2 size in 10 patients were randomly selected to examine the hair survival OVer 8 months post-operation.Tissue expansion was performed by firstly positioning a tissue expander in the subgaleal pocket of scalp and subsequently inflating it with normal saline in 5-7 days intervals for about 3 months. Thereafter, an expanded scalp flap was designed, with the combination of advancement and rotation flap transplantation principles, to be transferred to the recipient site after lesion removal. Results In patients with hair-grafting technique, hair density reached 60 -80 hairs/cm2 per session. Over an 8-month follow-up, the grafted hairs grew well with 98% hair survival. Forty-five patients with 20% -30% scalp hair loss achieved very satisfactory results with only one session while 91 patients with 31% -50% hair loss needed a second session to improve their appearance. In those with tissue expansion, 56 patients with 20% -50% scalp loss were managed in one session and 26 patients with 51% -75% scalp loss needed secondary tissue expansion. All of the expanded hair-bearing flaps survived well with only minimal complications (9. 8% ) of injeetive dome explosion in 3, slight skull depression in 2, seroma in 5 and early infection in 1. Condusions Either hair grafting or tissue expansion is proven as a safe and effective technique for restoring large eicatrical scalp alopecia. The former technique could be of benefits to simple, mini-surgical procedure, natural hair growth and good for lesion size under 30%. And the latter technique shows wider clinical indications, especially those with unstable scar, skull defect under lesion and lesion over 50%. But the obvious drawbacks are multi-clinic visits, disfigurement and long-term procedures.

Objective To validate whether STATl participated in the process of CTGF-induced proliferation and differentiation of human hypertrophic scar fibmblast(hHSF)on the basis of previous researches.Methods hHSF was co-cul6vated with 6 patients'hypertrophic scar specimens.Electrophoretic mobility 8hilt assay(EMSA)was used to verify binding ability between DNA and STATl with the stimulus of difflerent concentrations of CTGF(0,5,7.5,10,15 ng/ml) at 45th min and the stimulus of 10ng/ml CTGF at different phase point(0,10,20,30,45,60,90,and 120 min).We divided cells into CTGF group, STATI ASODN group.STAT1 ASODN+CTGF group and control group.And MTT was used to detect the proliferation of hHSF on daysl,2 and 3,and RT-PCR to detect α-smouth muscle actin mRNA to follow the differentiation.Results EMSA showed that the binding ability between STAT1 and DNA depended on the concentration of CTGF and peaked with the stimulation of 10 ng/ml CTGF.And at the same time,it peaked at 45-60 min wiht 10 ng/ml CTGF.MTT showed that cellular proliferation of CTGF group wag much higher than that of control group(all P<0.05).And those of STAT1 ASODN group and STATI ASODN+CTGF group were much lower than those of control group and CTGF group(all P<0.05).RT-PCR showed that differentiation activation from fibroblast to myofibwblast of CTGF group,STATI ASODN group,STAT1 ASODN+CTGF group and control group were 0.78±0.08,0.38±0.09,0.76±0.10,and 0.40±0.12,respectively. Differentiation activation of STAT1 ASODN group and control group were much lower than those of CTGF group and STAT1 ASODN + CTGF group ( all P < 0. 05 ) Conclusions STAT1 ASODN is important in the proliferating process of hHSF and it blocks the stimulation of CTGF on hHSF proliferation.The above result revealed that STAT1 participates in the process of hHSF proliferation induced by connective tissue growth factor.

Objective To investigate the effects of Notch signaling on scars in a rabbit ear model of hypertrophic scarring.Methods The hypertrophic scar of rabbit ears was reproduced.The lefl rabbit ear wounds as the N-[N-(3,5-dilluorophenacetyl-L-alanyl)]-(S)-phenylglycine t-butyl ester(DAPT)treated group were treated intradernudly with the γ-serelame inhibitor DAPT to inhibit the activation of Notch on day 1,3,7 and 14.The right ears as the control group were treated with normal saline at the samw time points.Experimental and control wounds were harvested on day 14,21,28 and 35 post-wounding,and then examined histologieally to quarttify hypertrophic index and fibroblasts.The expression of epidermal differentiation markers-keratin 19(K19),keratin 19(K14),involucrin and Notch downstream molecules-P21,P63 were examined and analyzed with inununohistochemical staining.Results On day 21,28 and 35post-wounding,both hypertrophic index(1.93±0.32,1.82 4±0.36,and 1.79±0.25,respectively)and number of fibroblasts[(4.08±0.88),(3.30±0.53),and(3.19±0.73)×103/mm2,respectively]in the DAPT treated group were significantly lower than those ofthe control group[2.56±O.29,2.61±0.30,2.58±0.39.and(5.45±0.99),(4.80±1.13),(4.43±1.17)× 103/mm2,all P<0.01)].The expression of K19,K14 and P63 increased in the DAPT treated group(28.6%±5.7%,53.1%±4.5%,and 57.0%±5.8%,respectively)as compared with the control group(10.1%±2.8%,30.8%4±4.9%,and 16. 5%±2. 2%, all P <0.01 ) on day 14 post-wounding, while the expression of involucrin and P21 decreased in the DAPT-treated group ( 12. 3% ±1.9% and 11.0% ±1.7% respectively) as compared with the control group (29.3%±4.6% and 44.3%±3.5%, both P<0.01). Conclusion Blocking of Notch signaling will inhibit the over-differentiation of scar epidermis, and thereby suppress the proliferation of hypertrophic sears in rabbit ears.

Objective To discuss the potential mechanisms and the role of adrenaliris with extensive hemorrhage and necrosis in acute critical infectious diseases.Methods Forensic autopsy cases from 1998 to 2007 were retrospectively analyzed.And 10 cases of them were involved in the primary and critical infecfion with acute infiammation,hemorrhage and necrosis of adrenal glands.Results Clinical diagnoses of 10 Cases were respiratory tract infection(n=6),septic shock(n=2),acute esophagitis(n=1),and undefined causes(n=1),and the quickly aggravatinng conditions resulted in death ultimately.However,systematic autopsy confirmed that 7 cages died of acute lymphocytic pancarditis,2 cases acute and lymphocytic interstitial pneumonia and 1 cKse epidemic cerebrospinal meningitis with septicemia.And severe complications of all cases included hemorrhagic necmtizing adrenalitis,systemic inflammatory response syndrome and multiple organ failure(MOF).Conclusions Hemorrhagic necrotizing adrenalitis should be considered as one of the important pathological changes in fatal MOF resulting from severe infection and trauma etc.More attention should be paid in clinical treatment.

Objective To evaluate the side effects of non-steroidal anti-inflammatory drugs (NSAID)on gastric mucosa and to study the preventive effects of teprenone in patients.Methods A total of 108 patients taking NSAID for more than 3 months with no infection of Helicobacter pylori(Hp)were recruited. All patients were Bcrleened by endoscopy and their upper gastrointestinal symptores were evaluated. Then,after excluding 16 patients with ulcer8,92 patients were randomly divided into intervention group with teprenone and control group.After follow-up for 3 months,the patients were re-screened by endoscopy and their upper gastrointestinal symptoms re-evaluated.Specimens of gastric mne06a were studied by PAS staining,and cyclooxygenase(COX)level were evaluated by immunohistuchemical technique.Results Among the patients taking NSAIDs,erosion occurred in 48(44.4%)patients while 16(14.8%)were found with peptic ulcers.The damages improved significantly(Z=-4.96,P=0.000)in the intervention group with teprenone(n=45)as compared with the control group(n=47)after follow-up for 3 months.Both COX-1 Jevel[31.1%(14/45)vs 6.7%(3/45),P=0.003] and mucous thickness [66.7%(30/45)vs 13.3%(6/45),P=0.000]also increased in the intervention group as compared with the control group.No significant difierenee was found in COX-2 level between these two groups[28.9%(13/45)vs 31.1%(14/45),P=0.82].Conclusion Long-term use of NSAID caused severe damages to gastroduodenal mucosa;teprenone improved NSAID-related gastric side effects and increased COX-1 level and mucous thickness.

Objective To investigate insulin secretion function and insulin resistance in Chinese newly diagnosed type 2 diabetes ( obese and non-obese patients) in order to provide evidence for clinical treatment. Methods A total of 408 newly diagnosed type 2 diabetes and 40 normal controls were recruited.Height and weight were measure& Insulin and glucose of 0, 30, 60 and 120 rain during oral glucose tolerance test were examined. The patients with fasting glucose level greater than 8.3 mmol/L were treated with gliclazide for 1-3 months. After normalization of plasma glucose levels for more than 2 weeks, this medication was withdrawn for 48 hours then OGTT repeated to assess IR and IS. Results The patients were divided into four groups based on fasting plasma glucose ( DMI : FBG <6. 9 mmol/L; DM2: 6. 9 mmol/L≤FBG < 8. 3 mmol/L; DM3 : 8. 3 mmol/L≤FBG < 9. 7 mmol/L; DM4 : FBG≥9. 7 mmol/L). Each group was further stratified to subgroups by cut-off point of BMI = 24 kg/m2. The insulin sensitivity and insulin secretion function were compared between the subgroups. (1) True insulin level in BMI≥24 (FBG < 6. 9mmol/L) subgroups were higher than controls ( 3.5±0. 5 vs 3. 2 ±0. 6 natural logarithm) ( P < 0. 05 ).(2) In BMI≥24 subgroups, the insulin sensitivity was even worse than BMI <24 groups, but the insulin secretion function was better at the same FBG level. (3) After intervention, the change of insulin sensitivity in Bmi <24 groups was better than BMI≥24 groups ( -4. 7±0. 9 vs -5.5±1.4 natural logarithm) (P <0. 05) ; but the change of insulin secretion function in BMI < 24 groups was worse. Conclusions ( 1 ) In newly diagnostic type 2 diabetes, insulin sensitivity and insulin secretion function decreased with the rising FBG, but they were different between the obese and non-ubese groups. (2) Insulin secretion function recovered better in the obese group when ameliorating glucose toxicity.

Objective To expiore the association between gene mutation of transcription factor Nkx.5 and Chinese patients with congenital heart disease(CHD).Methods Polymerase chain reaction (PCR)and DNA sequencing were used to check 99 CHD patients and 90 normal control subjects from Zhong Da HospitaJ of Southeast University.After arnplifying exons 1 of Nkx2.5 gene by PCR.We purified the PCR products and conducted the sequencing reaction,analyzed the mutation screening of exon 1 of Nkx2.5,investigated whether or not the Nkx2.5 WaS related with CHD in Chinese population.Resnlts One mumfion (A239G)in exon 1 of Nkx2.5 was identified in 3/90 normal control subjects and 12/99 CHD patients,including 3/24 with VSD,7/35 with ASD,1/13 with PS and 1/21 with PDA.Conclusion There are some associations between Nkx2.5 gene mutation and occurrence of eongenital heart disease in Chinese people.

Objective To explore the relationship between expression and regulation of megalin in gallbladder mucosa and cholesterel gallstone disease.Methods Gallbladder mucosa,gallbladderr wall,bile and gallstone were collected from 29 patients with cholesterol gallstone disease(GS)and 12 patients with gallstone free(GSF).Lipids of bile and stone were meagured by kits.Expression of megalin and eubilin was analyzed by real-time PCR GBC-SD cell line was treated with T0901317,9-cis ratinoic acid,chenodeoxycholic acid(CDCA)and agonists of LXR,RXR,FXR respectively.Gene expressions were detected.Results Biliary eholesterol % molar and CSI increased significantly in GS group[(7.98±0.44)mol% vs(4.87±0.39)mol%,P<0.01].Megalin expression in GS group was significantly higher than that in GSF group(P<0.05)and cubilin expression was similar between two groups.In vitro experiments showed that CDCA markedly increaged the expression of megalin.Conclusions The increaged expression of megalin may help to increase cholesterol uptake in galibladder and play a compensative role in GS.FXR may participate in the transcriptional regulation of megalin.

Objective To study the eorrelation between the expression of matrix metalloproteinase (MMP)-2,MMP-9,vascular endothelial growth factor(VEGF)and vasculogenic mimicry(VM)in gastrointestinal stromal tumors(GIST).Methods The immunohistochemical staining indiees(SI)0f MMP-2,MMP-9,VEGF were assessed on specimens of 84 human cases with GIST(21 VM-positive cases).Gelatin zymography analysis of MMP-2 and MMP-9 activities were performed on another 42 human cases of GIST with fresh tissue(22 VM-positive cases).Results The staining indices(SI)0f MMP-2 and MMP-9 were higher in the VM-positive group(4.10±2.05 and 3.43±1.89 respectively)than in the VM-negative group(2.98±1.97 and 2.38±1.84 respectively,both P<0.05);there was no statistical difierence in the SI of、VEGF between VM-positive and VM-negative groups.Gelatin zymography analysis showed that the activities of MMP-2 and MMP-9 were significantly higher in the VM-positive group(3.62±3.95 and4.77±5.29 respectively)than in the VM-negative group(1.26±1.2l and 2.11±1.54 respectively.both P<0.05).Conclusion The expressions of MMP-2 and MMP-9 correlate with VM formation in GIST.

Objective To investigate the effects of Shenmai injection containing active principles of Ginseng and ophiopogon root on the expression of hypoxia—inducible factor 1-α(HIF-1 α)in brain after hypoxic-ischernic brain damage(HIBD)．Methods 108 neonatal SD rats were randomly divided into 2 equal groups：①Shenmai group(Group SM)，undergoing ligation of the right common carotid artery to establish HIBD models,breathing immediately a mixed gas with 8％oxygen and 92％nitrogen for 2 h to cause HI insuh,and then injected intraperitoneally with Shenmal injection 10 ms／kg once a day for 7 d，and ②normal saline group(Group NS)undergoing ligation ofthe right common carotid artery to establish HIBD medels,breathing immediately a mixed gas with 8％oxygen and 92％nitrogen for 2 h，and then injected intraperitoneally with NS 10 ms／ks once a day for 7 d Another 54 neonatal rats underwent sham operation but did not undergo hypoxia as control group(Group C)，2，12，and 24 h，and 3，7，and 14 d after HI insult 9 rats from each group were killed with their right hippecampal tissues taken out Flow cytometry was used to examine the apoptotie rate of the hippocampal neurons．RT-PCR was used to detect the mRNA expression of HIF-1α. Results(1)The apoptesis rate of the-right hippoeampaI tissues began increase 2 h after HI insuh．peaked 24 h after HI，then gradually decreased，and almost returned to the original levels 14 d after HI．There was no significant differences in apoptosis rates 14 d after HI among the 3 groups(all P>0．05)．The neuron apoptosis rates 12 h，24 h，3 d，and 7 d after Hl of Group SM were all significantly Iower than those ofGroup NS(e．g 24 h：(11．95±1．13)％ vs(16．80±1．44)％，all P<0．05)．(2)The HIF-1α mRNA expression level in right brain began to increase 2 h after HI，peaked 24 h after HI，then gradually decreases，and returned to the original level 14 d after HI in both Group SM and Group NS；the HIF-1α mRNA expression in right brain 12 h，24 h，3 d，and 7 d after HI of Group SM were all significantly higher than those of Group NS ( e.g 24 h: ( 44. 32±4.03 ) % vs ( 35.63±3.73 ) %, all P < 0.05 ).Conclusion The HIF-1α mRNA expression in brain tissue is up-regulated after HI insult. Shenmai injection helps increase the mRNA expression of HIF-1α in brain and reduces the apoptosis of hippocampus neurons after HI insult.

Objective To detect the effect of functional status and immunophenotype of dendritic cell modified witll growth hormone(GH)from cord blood.Methods Monocyte wa8 isolated from fresh cord blood.DCs were induced with IL-4 & GM-CSF and medified with different concentrations of GH.The morphology,count and growth status of DC were observed appearance.And the immunophenotype of DC was detected with flow cytometry.IL-12 in supernatant wa8 measured with ELISA.The inununo-activity was determined with autologous mixed lymphocyte reaction.Results Mature DC were induced from adherent lymphomonoeyte under the action of IL-4.GM-CSF and GH of different densities(5,25,100 ng/ml).GH increaes the positive expression of CD83(31%±4%,34%±4%,34%±5%),CD80(26%±4%,29%±6%,30%±6%)and HIA-DR(45%±8%,49%±6%,50%±5%)(P<0.05),enhanced theability of secreting IL-12(P<0.05)and boosted significantly the immuno-activation of DC to autologous lymphocyte(P<0.05).Conclusion GH could promotethe differentiation,maturationand expression of co-stimulatory molecules,increase the IL-12 production in DC and enhance the ability of DC to stimulate the proliferation of lymphocyte within a certain dose range.

Objective To explore Sift innovative method for preparing a whole-liver reconstruct scaffold with intact three-dimensional geometry, vasculature and bile duct by decellularization technology.Methods The portal vein was annulated and perfused sequenfially with 1% Triton X-100 and 1% SDS for about 4 h,and then was perfused with phosphate buffered saline to dilute SDS residue.The retained stmcRlre was evaluated by histological analyses,including macroscopic,Hematoxylin-Eosin staining,Masson's trichrorne staining,orcein staining and SEM.The liquid polymer preparation 8%-10%,which was made of chlorinated poly vinyl chlofide(CPVC as solute),acetone(as solvent)and pigment,was injected into portal vein and bile duct to demonstrate tIle integrity of the portal vein and bile duct. The scaffold was cut into slices with the thickness of about 50μm and co-cultured with C3A cell line.Results Macroscopic examination showed that the deedlularized liver WS8 transparent and intrahepatic Gli8son's system could be observed.H&E staining of slices of decellularized liver demonstrated no intact cells or nuclei existect Masson trichrome staining revealed collagen retained.Orcein staining showed that there were elastic fibers.SEM showed the network of ECM Was intacL C3A-to-scaffold co-culture revealed the scaffold of a good biocompatibility.Conclusion Peffusion with detergents throuSh portal vein for liver decellularization is an efllcient method to obtain a completely whole-1iver scaffold for hepatic organ reconstruction.

：Objective Theoxalate-degradation genes of Oxalobacter formigenes(Ox.F)-frc and oxc genes were clonedand transfected into murine intestinal stem eell population to make the latter acquireoxalate-degradation function.Methotis(1)The dicistronic eukaryotic expressionvector,co-expressing OXC and frc genes,pIRES-oxc-frc was constructed.(2)The murineintestinal stem cell population wag isolared and cultured,and the function of cellulargrowth and differentiation identified.(3)The cells were transfected withpIRES-oxc-frc.After selection by G418,the function of cellular growth and differentiatielland the expression of objective genes were identified.(4)The concentration of oxalate inthe culture medium growing the transgenic cells wag determined by ion chromatography toexplore the oxalate-degradation function of cells.Results Ox.F could be isolated andcultured from the feces of Chinese people.Compared with foreign reports,a certainmorphologic variation of Ox.F existed.But OXC and frc genes showed a high homology withthe sequence reported in GenBank.The recombinant plagmid containing OXC and frc genescould be successfully transfected into the murine intestinal stem coll population.Theexpression of objeetive genes wag normal.The concentration of oxalate in the culture fluidof ransgenic intestinal stem cell population[(2.48±0.03)g/L]was lower than those of thenormal group[(2.69±0.01)g/L]and the control group[(2.69±0.01)g/L,P<0.01].ConclusionsBoth oxc and frc genes could be transfected into the murine intestinal stem cellpopulation,and the cells could be given oxalate-degrading function.The gene ofprokaryecyte could be introduced into the eukaryocyte for u successful expression.

Skin wound healing is a very delicate and complicated process. Excessive healing will lead to the formation of pathological scars, which are manifested as abnormal cytokine secretion, enhanced local inflammatory response, abnormal proliferation and apoptosis of fibroblasts, accumulation of extracellular matrix, etc. The proliferation of tissues will affect the appearance and even organ function of patients. Therefore, the prevention and treatment of scar formation after skin wound is an important issue in the field of wound healing research. With the deepening of research on wound healing and pathological scar formation mechanism, more and more molecules have been found to be related to skin scar formation.

Examination is a measure of students 'academic achievement and teachers' teaching level, and it is an important link in education.The traditional examination method basically adopts written examination, which basically examines theoretical knowledge, and it is difficult to investigate practical knowledge.This examination method is easy to urge students to read dead and memorize to obtain high scores, but neglect the pursuit of practical ability, and become students with "high scores and low ability". The unified examination of university enrollment reflects this situation, so the middle school education in our country is often regarded as "examination-oriented education" and is criticized.

Acute exacerbation of asthma is associated with airway inflammation, especially eosinophils. In necropsy patients who died of acute severe asthma, infiltration of activated eosinophils in airway mucosa can be observed. At the same time, the control of eosinophils airway inflammation is associated with the reduction of the number of acute attacks. Mepolizumab is a monoclonal antibody to interleukin 5 (IL-5), which can selectively inhibit eosinophils airway inflammation, so it is presumed to be effective for acute asthma attacks.

Smoking is one of the leading causes of early death in Americans,and about 438,000 people die from smoking every year.70% of smokers have the intention to quit,but only 2% ~3% of people quit successfully every year.Although smoking cessation programs and drug treatment can improve the success rate,the participation rate of smokers in these programs and the use rate of therapeutic drugs are very low.Dr. Volpp and others from the University of Pennsylvania conducted a large-scale randomized controlled study on the role of financial incentives on smoking cessation,which showed that financial incentives can help long-term smoking cessation.The results of the study were published in the February 2009 issue of the New England Journal of Medicine (N Engl J Med, 2009, 360:699-709).

At present, there is no consensus on whether ozone increases the mortality associated with environmental pollution. Dr. Jerrett of the University of California, USA, and others studied whether long-term exposure to ozone increases the mortality caused by cardiopulmonary diseases, especially those caused by respiratory diseases. The results were published in March 2009<New England Journal of Medicine >(N Engl J Med, 2009, 360: 1085-1095).