Objective To detect the expression pattern of microRNA in A549 cells treated by lipopolysaccharide, study the expression of miRNA-1247-3P in A549 cells under LPS treatment and explore the possible mechanism of miRNA-1247-3P in A549 cells under LPS treatment. Methods A549 cells were divided into experimental and control groups. Immunocytochemical method and RT-PCR were used to detect the changes of SP-A and SP-C. The expression of miRNAs were detected using miRNAs array in different groups. The key miR-1247-3P was collected to detect the changes of miR-1247-3P in all groups using quantitative real-time polymerase chain reaction. Results Compared with control group, the expressions of SP-A and SP-C were significantly decreased in the experimental groups (P < 0.05). MiRNA array showed that 31 miRNAs were up-regulated and 3 miRNAs were down-regulated. Compared with control group, the expression of miR-1247-3P was significantly increased in the experimental groups (P < 0.05). Conclusion The increased expression of miR-1247-3P may play an important role in the pathogenesis of ARDS.

Objective To investigate the effect of spinal cord ischemia/reperfusion injury on hindlimb dysfunction in rabbits.Methods Twenty-eight health adult rabbits were distributed into normal control group (n =4) and model group (n =24) according to the random number table.The modelof spinal cord ischemia/reperfusion injury was established by selective occlusion of lumbar arteries.The model group were submitted to ischemia for 30 min (Group Ⅰ),60 min (Group Ⅱ) and 90 min (Group Ⅲ) before the reperfusion with 8 rabbits each.Jacobs score,Reuters score and Rivlin inclined plane test were used to evaluate the hindlimb function in each Group at days 1,3 and 7 after reperfusion.Changes in nerve conduction function in each group were observed using the cortical somatosensory evoked potential (CSEP).Results At days 1,3 and 7,the paraplegia rates in group Ⅰ were 50％,38％ and 38％ respectively,in Group Ⅱ were 75％,88％ and 100％,and in Group Ⅲ were all 100％.Paraplegia rate differed significantly among the three groups at 1 d and 3 d (P ＜ 0.01).Paraplegia rate differed significantly in Groups Ⅱ and Ⅲ when compared to that in Group Ⅰ at 7 d (P ＜ 0.01),but there was no significant difference between Groups Ⅱ and Ⅲ (P ＞ 0.05).With the prolongation of reperfusion,the Reuters score in Group Ⅰ dropped but not differed from that in control group (P ＞ 0.05);the Reuters score in Groups Ⅱ and Ⅲ increased and differed from that the control group (P ＜0.01),but the difference between Groups Ⅱ and Ⅲ was insignificant (P ＞ 0.05).Critical angle and obstacle rate of the inclined plane in control group were (68.4 ± 3.0)° and 0％.One day after reperfusion,critical angles of the inclined plane in Groups Ⅰ,Ⅱ and Ⅲ were (58.8 ± 4.1) °,(38.5 ± 2.8) ° and (29.8 ± 1.8) °,and the obstacle rates were (14.5 ± 0.9) ％,(43.6 ± 2.4) ％ and (56.0 ± 2.9) ％.There were significant differences compared to control group (P ＜ 0.01).Slight decrease in critical angle of the inclined plane but a minor increase in the obstacle rate was detected in Groups Ⅱ and Ⅲ at 3 d and 7 d after reperfusion,and the differences were significant compared to control group (P ＜ 0.01).Three days after reperfusion,critical angle of the inclined plane raised and obstacle rate of the inclined plate fell in group Ⅰ,not significantly different from these in control group (P ＞ 0.05).Latencies of CSEP N1 and P1 waves in Group Ⅱ [(33.1 ± 1.8) ms and (58.6 ± 4.0) ms] were longer than these in control group [(23.7±0.5)msand (48.1±4.1)ms]andgroup Ⅰ [(26.2±0.7)ms and (50.2±4.2)ms] (P＜ 0.01) 7 days after reperfusion,but the differences between control group and Group Ⅰ were insignificant (P ＞ 0.05).While the CSEP wave disappeared in Group Ⅲ.Conclusions Severity of spinal cord inschemia/reperfusion injury is related to the duration of ischemia.Hindlimb dysfunction caused by ischemia/reperfusion injury is characterized mainly by spastic paraplegia.

BACKGROUND: Some studies in vitro have reported that there are CD30 positive T cells in immunological response of allogenic transplantation.OBJECTIVE: To detect the relationship between the level of serum CD30 (sCD30) and clinical rejection in the patients with or without kidney transplantation, and analyze the importance of sCD30 in the estimation of immune state, monitor of acute rejection, and judgment of prognosis. DESIGN, TIME AND SETTING: Clinical case analysis study was performed at Jiangsu People's Hospital between April 2004 and March 2007. PARTICIPANTS: 153 kidney transplantation cases comprising 103 males and 50 females, averagely aged 37 years. METHODS: 3 mL peripheral blood was obtained from recipients before transplantation (without immunosuppressive agent) and at 0, 1, 3, 5, 7, 14, 21, and 28 days. Serum was isolated from obtained blood and placed at -20 ℃. Soluble CD30 levels were detected using CD30 cytokine ELISA kit supplied by BenderMedSystems. MAIN OUTCOME MEASURE: The relation between the soluble CD30 levels and rejection prior to and following transplantation.RESULTS: There was a significant relation in the sCD30 level between the patients with (n=17) and without acute rejection (n=136). The CD30 levels were 113.2 U/mL in the rejection group and 83.2 U/mL in the non-injection group (P < 0.01). No significant difference was determined between both groups in 5 days following surgery (P > 0.05). Significant difference were detected between both groups from 5 days following surgery (P < 0.01). There was no relation between the soluble CD30 level and the time of rejection and release after kidney transplantation (P > 0.05). Receiver operating characteristic (ROC) curve demonstrated that soluble CD30 levels on day 5 post-transplantation could predict acute rejection (area under ROC curve: 0.850). Meanwhile, 100 U/mL was the optimal operational cut-off level to predict rejection (specificity: 85.0%; sensitivity: 83.6%). The patients with positive of soluble CD30 level showed a lower survival rate than those with negative CD30 level (P < 0.01). CONCLUSION: The soluble CD30 levels contributed to predictive the acute rejection and prognosis of kidney transplantation.

Objective To investigate the effects of antioxidant SS-31 on sepsis-induced acute lung injury (ALI)in a mouse model of sepsis.Methods Sepsis was induced in male mice by cecal liga-tion and puncture (CLP).Forty-eight adult male mice (C57BL/6,weight 25-32 g)were equally as-signed to the sham+vehicle group (group A),sham+SS-31 group (group B),CLP+vehicle group (group C),or the CLP+SS-31 group (group D).At 0 or 5 h after CLP or sham operation,mice re-ceived an intraperitoneal injection of SS-31 (5 mg/kg of body weight)or the same volume of normal saline.Pulmonary tumor necrosis factor alpha (TNF-α),interleukin (IL)-6,IL-10,malondialdehyde (MDA),superoxide dismutase activities (SOD),myeloperoxidase activities (MPO ),wet-to-dry weight ratio (W/D),reactive oxygen species (ROS),ATP,NF-κB p65,inducible nitric oxide syn-thase (iNOS),and histological scores were assessed 24 h after operation.Results Pneumonia,edema were significantly heavier in group C than in group A (P <0.05).Lung congestion,inflammatory cell infiltration,alveolar wall edema was significantly less in group D than in group C (P <0.05).Pulmo-nary histological scores,IL-6,MDA,MPO,W/D,ROS,NF-κB p65 and iNOS significantly in-creased,while ATP levels decreased in group C when compared with group A (P <0.05).However, SS-31 treatment significantly reversed these parameters when compared with the group C (P <0.05). No difference was observed between the group A and group B.There was no difference of TNF-α,IL-10,and SOD among the four groups.Conclusion SS-31 improves sepsis-induced ALI in a mouse model probably by down-regulating the oxidative stress and inflammation in of sepsis-induced ALI.

Objective To investigate the chemical properties of superficial cardiac neurons. Methods By means of immunohistochemical ABC technique,the study was performed concerning the distribution of calcitonin gene-related peptide(CGRP) and substance P(SP) in the canine main cardiac superficial plexus. Results CGRP-immunoreactive(IR) neurons were found in every plexus,but SP-IR neurons could be observed only in dorsal atria plexus(DAP),inter atria plexus(IAP) and aorta-pulmonary plexus(A-PP).The shape and size of CGRP-IR and SP-IR neurons were similar.The comparative study on atria and ventricles indicated that CGRP-IR and SP-IR neurons in atria were more than those in ventricles.Numerous CGRP-IR,SP-IR nerve fibers could be observed in each fat pats and intermyocardiocytes.These nerve fibers were usually situated near blood vessels,or were attached to vessel wall.Somewhat CGRP-IR and SP-IR nerve fibers were connected with myocardiocytes in some regions.Conclusion The results indicated that actually existed two peptides in canine cardiac superficial plexus.These implied that the regulations of the two kinds of peptidergic neurons to atria and ventricle were different.The two kinds of neurons mentioned above were likely to perform different or similar functions in canine heart.CGRP and SP could possibly modulate the activities of myocardiocytes and vessels of heart directly.

AIM: To prepare and identify mouse polyclonal antibody against protein Hlb, which is the variant of major subunit of human ASGPR. METHODS: Hlb specific peptide was synthesized and coupled with keyhole limpet hemocyanin (KLH) for immunization. Then H1b-KLH conjugation was injected into mouse subcutaneously to produce polyclonal antibody. ELISA assay was used to detect the titer of the antibody. Antibody was also identified by Western blot and immunohistochemistry assays. RESULTS: Mouse antibody against Hlb was prepared after injection of H1bKLH conjugation. The titer of H1b antibody was about 1:10~5.Western blot confirmed its high specificity. This antibody could also be used for immunohistochemistry analysis. CONCLUSION: The successful preparation of the polyclonal antibody against protein H1b, which can discriminate the two variants of the major subunit of ASGPR with high specificity, will provide an efficient reagent for further study of the physiologic functions of H1b and its role in the pathogenesis of human disease.

Objective To evaluate the therapeutic effects and to summarize the clinical experience of benign prostatic hypertrophy(BPH) with inguinal hernia in the elderly 70-89 years. Methods Clinical data of 32 patients 70-89 years old simultaneously undergone transurethral resection of prostate (TURP) and plug-mesh tension-free hernia repair from July 2000 to May 2005 were retrospectively analyzed and followed up. Results The average operating time was (85.0?12.8)minutes, the average blood loss was (90.0? 18.7 )ml. No postoperative death or life threatening complications were revealed. By 7-40 months of following up reported that there were no recurrence of hernia as well as no incontinence and urethral stenosis or other complications. IPSS,maximal flow rate and residual urine were evidently improved after operation. Conclusion Combined TURP and plug-mesh tension-free hernia repair is a safe and effective procedure for the elderly patients.

Hepatitis B virus surface antigen(HBsAg),a specific antigen on the membrane of Hepatitis B virus (HBV)-infected cells,provides a perfect target for therapeutic drugs.The development of reagents with high affinity and specificity to the HBsAg is of great significance to the early-stage diagnosis and treatment of HBV infection.Herein,we report the selection of RNA aptamers that can specifically bind to HBsAg protein and HBsAg-positive hepatocytes.One high affinity aptamer,HBs-A22,was isolated from an initial 115 mer library of ～1.1×1015 random-sequence RNA molecules using the SELEX procedure.The selected aptamer HBs-A22 bound specifically to hepatoma cell line HepG2.2.15 that expresses HBsAg but did not bind to HBsAg-devoid HepG2 cells.This is the first reported RNA aptamer which could bind to a HBV specific antigen.This newly isolated aptamer could be modified to deliver imaging,diagnostic,and therapeutic agents targeted at HBV-infected cells.

OBJECTIVETo study the effect of tongfei mixture (TFM, a Chinese recipe mainly consisted of angelica and rehmannia root) on nocturnal hypoxia in patients with chronic obstructive pulmonary disease (COPD).

METHODSSixty patients with COPD of remission phase were randomly divided into 3 groups, 20 in each group. Group A was the control group; Group B, the group simply treated with oxygen; Group C, treated with oxygen and TFM. Changes of pulmonary function, diaphragm muscle mobility (DMM), 6 min walk distance (6MWD), morning arterial blood gas, nocturnal lowest oxygen saturation (LSaO2), mean blood oxygen saturation (MSaO2), the percentage of saturation lower than 90% time account for total sleeping time (SLT90%) and ultrasonocardiogram before and after treatment were observed.

RESULTSLevels of LSaO2, MSaO2 and SLT90% in Groups B and C were significantly higher than those in Group A (P<0.05, P<0.01). The lowering of PaCO2 in Group C was more significant than that in Group B (P<0.05). The mPAP level in Group C was lower, FEV1, 6MWD and DMM were improved than those in Group A and B, showing significant difference (P<0.05).

CONCLUSIONCombined use of oxygen therapy and TFM could not only improve the nocturnal hypoxia, but also lower PaCO2. TFM is an important supplement of oxygen therapy.

Aged ; Blood Gas Analysis ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Hypoxia ; drug therapy ; etiology ; Lung Diseases, Obstructive ; complications ; drug therapy ; Male ; Middle Aged ; Oxygen Inhalation Therapy ; Phytotherapy ; Sleep ; physiology

To better understand the effect of a new split variant of human asialoglycoprotein receptor (ASGPR H1b) on ASGPR ligands' binding ability, we established a functional cell line which expresses ASGPR. The full lengths of ASGPRH1a and H2c fragments from human liver were amplified by reverse transcript PCR (RT-PCR) and inserted into eukaryotic expression vector pIRES2EGFP, pCDNA3.1 (Zeo+) respectively. The recombinants were co-transfected into HeLa cells. After selection by using Neocin and Zeocin, a stably transfected cell line was established, which was designated 4-1-6. The transcription and expression of ASGPRH1a and H2c in 4-1-6 were confirmed by RT-PCR, Western blotting and immunofluorescence. The endocytosis function of the artificial "ASGPR" on the surface of 4-1-6 was tested by FACS. It was found that the cell line 4-1-6 could bind ASGPR natural ligand molecular asialo-orosomucoid (ASOR). After the eukaryotic plasmid H1b/pCDNA3.1 (neo) was transfected into cell line 4-1-6, H1b did not down-regulate the ligand binding ability of ASGPR. The eukaryotic expression plasmid H1b/pcDNA3.1 (neo) and H2c/pcDNA3.1 (neo) were co-transfected transiently into Hela cell. Neither single H1b nor H1b and H2c could bind ASOR. In conclusion, a functional cell line of human asialoglycoprotein receptor (ASGPR) which expresses both H1a and H2c stably was established. The new split variant H1b has no effect on ASGPR binding to ASOR. ASGPRH1b alone can't bind to ASOR, it yet can't form functional complex with ASGPRH2c.