Objective To explore whether anoxia can induce expression changes in connective tissue growth factor(CTGF)in renal tubular epithelial cells(TECs)and epithelial-mesenchymal transition of TECs.Methods Rat renal TECs(NRK-52E)anoxia models were established.NRK-52E cells were exposed to anoxia for 4 h.The real-time RT-PCR,Western blotting,immunohistochemical staining were used to detect the expression of CTGF at 6,12,24,48,and 72 h in NRK-52E cells.Morphological changes and cytoskeleton remodeling in NRK-52E cells under anoxia were examined by a laser confocal microscope and BODIPYFL staining respectively.Results Under anoxia,NRK-52E cells became round,enlarged and cytoskeleton was remodeled.The expression levels of CTGF mRNA and protein were up-regulated at 6 h,reached their peak at 48 h:the expression of CTGF mRNA protein was 29.33±0.21 and 1.30±0.02 respectively.Under anoxia,NRK-52E cells underwent an epithelial-mesenchymal transition process,including cytoskeleton remodeling,and morphological changes.Conclusion Anoxia can change the expression of CTGF and other fibrosis-associated genes in NRK-52E cells,and CTGF played an important role in fibrosis process and epithelial-mesenchymal transition development in NRK-52E cells.

[Objective] To investigate the effects of REMl sleep deprivation (SD) on actions to investigate the efficacy of application of electro-acupuncture for treating sleep deprivation.[Methods]Thirty healthy SD rats randomly allocated to three groups (normal-control,model (A1),electro-treatment groups).The rats of model (A1)group and electro-treatment group were induced in male rats by housing them on small platforms overwater,normal-control group rats were on big platforms overwater.After electro-treatment for electro-treatment groups,the changes of normal status and Open Field Test were measured with all rats.[Results]Electro-acupuncture treatment can improve the actions.[Conclusion]Application of electro-acupuncture can produce a fighting effect on sleep deprivation.The effect may be related to promoting the nerval endocrine cytokine of electro-acupuncture.

A new dibenzocyclooctadiene lignan, renchangianin E (1) was isolated from the stems of Kadsura renchangiana. Its structure and stereochemistry were elucidated by spectroscopic methods, including 2D-NMR techniques.

Objective To investigate the effect ofulinastatin on severe heat-stroke with acute lung injury induced by severe heat stroke.Methods Thirty severe heat stroke patients were divided into conventional group (n=15) and ulinastatin group (n=15) randomly,with another 80 healthy adults serving as controls.The baseline data such as age,gender,onset period and APACHE Ⅱ scores were recorded and compared between the two groups on admission.Peripheral leucocyte counts,oxygenation index and Murray scores were determined on the 1st,3rd and 5th day.The concentration of inflammatory mediators in bronchoalveolar lavage fluid (BALF) and alveolar macrophage supernatant were detected by enzyme-linked immunosorbent assay (ELISA).Western blotting and real-time PCR were used to measure expression of triggering receptor-1 on myeloid cells (TREM-1) on alveolar macrophages.Furthermore,comparison was made in terms of the ventilation period,ICU stay time and mortality in 28 days between the two groups.Results No differences were found in age,gender,onset period and APACHE Ⅱ scores between the two groups (P＞0.05).Compared with the conventional group,peripheral leucocyte counts and Murray scores in the ulinastatin group significantly decreased on the 3rd and 5th day (P＜0.05,P＜0.01).But oxygenation index was higher in the ulinastatin group than in the conventional group (P＜0.05).The concentration of TNF-α and IL-6 in BALF was lower in the ulinastatin group than in the conventional group (on the 3rd day:P＜0.05,P＜0.01;on the 5th day:P＜0.01,P＜0.01).The concentration of TNF-α and IL-6 in alveolar macrophage supernatant was lower in the ulinastatin group than in the conventional group (on the 3rd day:-P＜0.05,P＜0.01;on the 5th day:P＜0.01,P＜0.05).The expression of TREM-1 protein on alveolar macrophages were lower in the ulinastatin group than in the conventional group (on the 3rd day P＜0.01;on the 5rd day P＜0.05).TREM-1 mRNA was lower in the ulinastatin group than in the conventional group (on the 3rd day:P＜0.05;on the 5th day:P＜0.05).Eventually,the treatment of ulinastatin shorten ventilation period and ICU stay time (P＜0.01,P＜0.05).Nonetheless,it failed to show any improvement in terms of the mortality during 28 days (P＞0.05).Conclusion Our study exhibited that ulinastatin had good effect on the heat stroke patients with acute lung injury and it helped reduce the inflammatory reaction of pulmonary tissues.The underlying mechanism of these effects might lie in its ability to reduce heat stroke-induced inflammatory secretion and expression of TREM-1 on alveolar macrophage.

Objective To study the association of glucagon like peptide 1 receptor (GLP1R) gene polymorphism with type 2 diabetes in Han population in Shanghai. Methods In the study, 360 type 2 diabetic patients and 313 normal control subjects were enrolled. Diabetic patients were further subdivided into insulin treated non obese patients (BMI28, 192 subjects). A single nucleotide polymorphism (SNP) rs 2268657 was genotyped in all the subjects enrolled in the study using allele specific real time PCR and its association with type 2 diabetes was examined. Results The frequencies of AA,AG, GG genotype incontrol group were0.086,0.447, 0.466 respectively, 0.155, 0.375, 0.470 in non obese diabetic patient group respectively, and 0.109, 0.500, 0.391 in obese diabetic patient group respectively. There was significant difference of the frequency of genotype AA between control group and non obese diabetic patient group (OR=1.939, P

Objective To is ol ate novel thyroid hormone-response genes, to study the characterizations of the ir expressions and to predict their possible functions in neonatal rats. Methods A neonatal rat model with congenital hypothyr oidism was established and cDNA fragments of novel thyroid hormone-response gen es from cerebral cortex of neonatal rats were obtained by fluorescence-labeled DD-PCR analysis, subcloning and sequencing. Complete cDNAs of novel thyroid hor mone-response genes were cloned by the techniques of electronic clone, RT-PCR and sequencing, their expressions regulated by thyroid hormone were confirmed b y Northern blot analysis, their distributions, transcription levels in different tissues and different brain areas were further observed by semiquantitative RT -PCR analysis, and their possible functions were postulated through bioinformat ic techniques. Results A novel complete cDNA of thyroid hormone-response protein-1 (TRP-1) gene is cloned. It is 973 bp in f ull-length (Gene Bank accession no. AF348365), and its transcription was enhanc ed in cerebral cortex in neonatal hypothyroidism rats. The expression of its mRN A was very extensive, but more abundant in brain. Its transcriptional level in d ifferent brain areas was not uniform, much higher in olfactory bulb. Its encodin g protein had some significant domains and motifs. Conclusion TRP-1 gene is a new thyroid hormone-response gene and may play an important role during normal brain development. Its abnormal expression may b e partially responsible for neurological defects in brain arising from thyroid h ormone deficiency during critical period for perinatal rats.

Objective To investigate the effects of different immunosuppressive agents on mesangial cell proliferation through a mesangial cell injury model in vitro. Methods Mesangial cell line (HBZY-1) in period of proliferation was cultured in vitro with cytochalasin B for 2 h, then HBZY-1 cells were divided into 5 groups: blank (control) group, cyclosporine A (CsA) group, Tacrolimus (Tac) group, mycophelonate mofetil (MMF) group and rapamycin (RAPA) group. Subsequently,the number of HBZY-1 cells at different time points was measured by using the professional image analysis software after treatment for 6, 12 and 24 h, respectively. Results Damaged HBZY-1 cells recovered in all groups. At 6 h, the number of HBZY-1 cells in Tac group was significantly more than that in control group (P＜0.05), but the difference had no significance between the other treatment groups and control group (P＞0. 05). At 12 h, there was no significant difference in of the number of HBZY-1 cells among the all groups (P＞0. 05). At 24 h, there was no significant difference in the cell number between MMF and control groups (P＞0. 05). CsA, Tac and RAPA resulted in HBZY-1 cell proliferation, and the cell number in CsA and Tac groups was significantly more than that in the other groups (P＜0. 05). As compared with the control group, the cell number in RAPA group was significantly increased (P＜0. 05). Conclusion CsA, Tac, MMF and RAPA contribute to recovery of damaged HBZY-1 cells, but CsA and Tac result in over-proliferation of HBZY-1 cells. RAPA and MMF can prevent HBZY-1 cells against over-proliferation, and MMF scarcely results in HBZY-1 cell proliferation.

Objective To study the association of single nucleotide polymorphism (SNP) of the protein tyrosine phosphatase-1B (PTP-1B) gene with type 2 diabetes mellitus and obesity in Chinese. Methods SNPs in the PTP-1B gene were detected by direct sequencing to PCR products, and the detected SNPs were genotyped in case-spouse samples with the technique of fluorescence real-time PCR. Results Totally 6 SNPs were found in PTP-1B gene. Three SNPs (I5/37 C→A,I6/82 A→G, I7/301 C→T) were in the intron regions and the other 3 (E8/45 C→T, E9/35 G→A, E10/372 G→A)in the exon regions. Among them, E9/35 G→A was a newly found mutation site. The A allele frequency of I5/37 C→A, T allele frequency of I7/301 C→T and G allele frequency of I6/82 A→G in type 2 diabetes were significantly higher than those in the normal spouse group (P

OBJECTIVESTo identify differences in gene expression in renal and visceral adipose tissue in type 2 diabetic rats using cDNA representational difference analysis (RDA) and to explore the molecular pathogenesis of type 2 diabetes and its chronic vascular complications.

METHODSA rat model of type 2 diabetes was generated by administration of a high fat and calorie diet combined with a low dose of streptozocin (STZ) injected into the tail vein. The difference bands were generated by cDNA representational difference analysis (cDNA RDA). The final difference products were ligated into the pUC-18 vector and sequenced. A bioformatics analysis was performed on the obtained expressed sequence tags (ESTs), and then the expression levels of known and novel genes were verified by semi-quantitative reverse transcription-PCR (RT-PCR). At the same time, full-length cDNA of a novel gene was cloned in silico.

RESULTSThe type 2 diabetic rats in this experiment experienced hyperglycemia, lipidemia, lower insulin sensitivity and normal body weight. We obtained 9 novel ESTs and 2 novel genes from renal tissue of rats and 6 novel ESTs and 1 known gene, the rat lipoprotein lipase (LPL) gene from their visceral adipose tissue. The 2 novel genes (RS91 and RS2) from the renal tissue were both very similar to serine (or cysteine) proteinase inhibitor, clade F and eukaryotic translation initiation factor 3 and subunit 5 (EIF-3 epsilon). The expression of both novel genes and the LPL gene were upregulated in renal and visceral adipose tissue of type 2 diabetic and fat-enriched rats. Full-length cDNA of the novel gene RS91 was cloned in silico.

CONCLUSIONS(1) The rat model of type 2 diabetes generated in this study was ideal because the disease in the animals closely mimicked type 2 diabetic patients. (2) cDNA RDA is a flexible, inexpensive, more accurate, sensitive and highly effective technique for identifying differences in gene expression. (3) Six novel ESTs and 1 known gene were obtained from rat visceral adipose tissue. The LPL gene was upregulated in adipose tissue of type 2 diabetic and fat-enriched rats, a result possibly related to the diabetic animals' high fat and calorie diet, lipidemia, insulin resistance (RI) and molecular pathogenesis. (4) Nine novel ESTs and 2 novel genes were obtained from the renal tissue of rat. We believe the 2 novel genes to be the serine proteinase inhibitors clade F and EIF-3 epsilon in rats. The upregulation of the 2 novel genes in renal tissue of type 2 diabetic rats and may have been related to their renal impairment.

Adipose Tissue ; metabolism ; Animals ; Cloning, Molecular ; Diabetes Mellitus, Type 2 ; metabolism ; Expressed Sequence Tags ; Gene Expression Profiling ; Kidney ; metabolism ; Male ; Oligonucleotide Array Sequence Analysis ; Plasmids ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Viscera

BACKGROUNDUp to now, locally advanced non-small cell lung cancer simutaneously involving carina, heart and great vessels is still regarded as contraindication for surgical treatment. However, the prognosis is very poor in these patients treated with chemotherapy and/or chemoradiotherapy. The aim of this study is to summarize the clinical experiences of carinoplasty combined with heart and great vessel plasty in the treatment of 84 patients with locally advanced non-small cell lung cancer involving carina, heart and great vessels or both in our hospital.

METHODSFrom March, 1988 to December, 2004, carinal resection and reconstruction combined with heart, great vessel plasty was performed in 84 patients with locally advanced non-small cell lung cancer involving carina, heart and great vessels simutaneously. The operative procedures in this series included as follows: (1) Right upper sleeve lobectomy combined with carinal resection and reconstruction, and right pulmonary artery sleeve angioplasty in 9 patients; (2) Right sleeve pneumonectomy combined with partial resection and reconstruction of left atrium, and superior vena cava resection and Gortex grafts in 3 cases; (3) Left upper sleeve lobectomy combined with carinoplasty, left pulmonary artery sleeve angioplasty and partial resection and reconstruction of left atrium in 3 cases; (4) Right upper sleeve lobectomy combined with carinoplasty, right pulmonary artery sleeve angioplasty and partial resection and reconstruction of left atrium in 10 cases; (5) Left upper sleeve lobectomy combined with carinoplasty and left pulmonary artery angioplasty in 9 cases; (6) Left upper sleeve lobectomy combined with carinoplasty, left pulmonary artery sleeve angioplasty and resection of the aorta arch sheath in 6 cases; (7) Right upper-middle sleeve lobectomy combined with carinoplasty and right pulmonary artery sleeve angioplasty in 3 cases; (8) Left upper sleeve lobectomy combined with carinoplasty, left pulmonary artery angioplasty, resection of the aorta arch sheath and partial resection and reconstruction of left artium in 8 cases; (9) Right upper sleeve lobectomy combined with carinoplasty, right pulmonary artery angioplasty and partial resection and reconstruction of left atrium in 4 cases; (10) Left sleeve pneumonectomy combined with partial resection and reconstruction of left atrium in 3 cases; (11) Right upper-middle sleeve lobectomy combined with carinoplasty, right pulmonary artery angioplasty and superior vena cava resection and reconstruction with Gortex grafts in 23 casese; (12) Right sleeve pneumonectomy combined with partial resection and reconstruction of left atrium in 1 case; (13) Right upper-middle sleeve lobectomy combined with carinoplasty, right pulmonary artery angioplasty and partial resection and reconstruction of left atrium in 1 case; (14) Right upper-middle sleeve lobectomy combined with carinoplasty, right pulmonary artery angioplasty and right inferior pulmonary vein sleeve resection and reconstruction in 1 case.

RESULTSThere were two operative death in this series. The operative mordality was 2.38%. A total of 32 patients had operative complications. The incidence of operative complications was 38.10%. The 1-, 3-, 5-and 10-year survival rate was 81.34%, 59.47%, 31.73% and 24.06% respectively.

CONCLUSIONS(1) It is feasible in technique that carinal resection and reconstruction combined with heart, great vessel plasty in the treatment of locally advanced non-small cell lung cancer involving carina, heart and great vessels simutaneously; (2) Multiple modality therapy based on carinal resection and reconstruction combined with heart and great vessel plasty can remarkably increase the survival rate, and improve the prognosis and quality of life in patients with locally advanced non-small cell lung cancer involving carina, heart and great vessels.