Objective To investigate the effect of propofol on LPS-induced TLR4 expression in rat alveolar type Ⅱ epithelial cells. Methods The primarily cultured alveolar type Ⅱ epithelial cells isolated from male rats were randomly assigned to one of 5 groups: group Ⅰ cells were incubated for 3 h without any additive (control) ; group Ⅱ cells were incubated with LPS 1 μg/ml for 3 h (LPS) ; group Ⅲ , Ⅳ , Ⅴ cells were incubated with LPS 1 μg/ml + propofol 25, 50 and 100 μmol/L respectively for 3 b (P1-3). TLR4 mRNA and TLR4 protein expression was detected by real time PCR and Western blot. TNF-α release amount was measured using ELISA. Results LPS significantly' increased TLR4 mRNA and protein expression in alveolar type Ⅱ epithelial cells as well as TNF-α release amount. Propefol at 50 and 100 μmol/L significantly inhibited LPS-imluced increase in TLR4 mRNA and protein expression and TNF-α release amount. Conclusion Propofol can dose-dependently inhibit LPS-induced inflammation in alveolar type Ⅱ epithelial ceils, through down-regnlation of TLR4 gene and protein expression.

The effect of the trace elements on retinopathy of prematurity (ROP) were studied. Thirty preterm infants who had potential high risk factors of ROP were selected as observation group and 18 normal infants as control groups. By using atom spectrophotometer, the contents of serum trace elements (Mg, Cu, Zn, Mn, Se) were measured and analyzed statistically. The contents of serum Zn, Cu and Se in observation group were 0.75+/-0.22, 0.41+/-0.20 and (134.07+/-71.57)x10(-3) mg/L respectively, and 0.55+/-0.12, 0.65+/-0.194 and (202.92+/-44.71)x10(-3) mg/L in control group respectively (P<0.01). The contents of Cu and Se were obviously lower and that of Zn higher in observation group than those in control group. The same results were obtained between the infants with ROP and controls (P<0.01). However, there was no significant difference in the contents of serum Mg and Mn between two groups (P>0.05). It was concluded that the contents of serum Cu and Se in preterm infants who had high risk factors of ROP were obviously lower than in the controls. The contents of serum Cu and Se in the ROP infants were also much lower while contents of Zn much higher. Attention should be paid to the detection of the trace elements in preterm infants in order to prevent the deficiencies of Cu and Se. Only in this way can we prevent the deficiencies of Cu and Se, so as to decrease the ROP risk factors and prevent the disease.
Copper/blood ; Infant, Premature/*blood ; Retinopathy of Prematurity/*blood ; Retinopathy of Prematurity/*prevention & control ; Risk Factors ; Selenium/blood ; Spectrophotometry, Atomic ; Trace Elements/*blood ; Zinc/blood

Adult ; Case-Control Studies ; Female ; Humans ; Male ; Nervous System Diseases ; etiology ; physiopathology ; Neural Conduction ; Organophosphate Poisoning ; Pesticides ; poisoning ; Young Adult

0.05;AGC R 1 to R 2 P 0.05.The invasive depth of the lesion corresponded to the degree of lymph node metastases.No relation was found between tumor size and the survival rate.The prognosis of type Ⅲ EGC and AGC with infiltration was poor.Conclusions: Biological behavior of gastric cancer can be used as a guide to operation type.We proposed that EGC should undergo subtotal gastrectomy plus R 1 or selective R 2 ,and AGC should undergo subtotal gastrectomy or near total gastrectomy plus R 2 or selective R 3 ,with a margin of at least 5 cm from the tumor.

Objective To construct cDNA entry library and cDNA expression library of Armillifer agkistrodontis (A.) nymphs and make a preliminary immunoscreening for the cDNA expression library.Methods The nymphs were collected from the Kunming mice infected experimentally with A.agkistrodontis eggs and the total RNA were extracted from the nymphs using TRIzol Reagent.After purifying the mRNA,the synthesized cDNAs were cloned into the donor vector pDONR222 by BP reaction of Gateway technology and the recombinants were transformed into the DH10B cells by electroporation,the cDNA entry library was obtained.Next,the expression vector pDEST17 was ligated with entry clones by LR reaction,and the recombinants were transformed into the BL21 (DE3) cells.Hence,the cDNA expression library was constructed.Then,the expression library was immunoscreened with the mixed sera of mice infected with A.agkistrodontis,and the insertions of positive clones were sequenced.After that,the open reading frame(ORF) of positive slone sequence,the homology of the screened genes and their encoded proteins were analyzed by Finder and BLAST (basic local alignment search tool) program of National Center of Biotechnology Information(NCBI),and the discovered new genes were submitted into the GenBank.Besides,the physico-chemical properties,secondary structure and B cell epitopes of encoded proteins were also analyzed by bioinformatics software.Results The average titer and total clones of the cDNA entry library were 1.45 × 105 CFU/ml(colony-forming unit,CFU) and 1.74 × 106 CFU,respectively,and the range of fragment length of the inserted cDNA was between 0.2-4.0 kb,with an average of 1.4 kb.The total clones of cDNA expression library were 1.00 × 105 CFU,and the fragment length of the inserted cDNA was between 0.3-2.2 kb,with an average of 1.0 kb.Five positive clones,coded S1,S5,A1,D1 and F1,respectively,were obtained through preliminary immunoscreening.The sequence and homology of the five positive clones were sequenced and analyzed by BLAST program.No significant similarities were found in pentastomida species,which meant that they were all novel genes of A.agkistrodontis.The gene sequences were submitted to GenBank,with the accession number from JQ180451 to JQ180455.Also,results obtained by bioinformatics software showed that the predictive encoding proteins were all potential to be valuable recombinant diagnostic antigens.Conclusions The cDNA library of A.agkistrodontis nymphs is successfully constructed,and five new genes of A.agkistrodontis are discovered.The establishment of cDNA library and the discovery of the new genes will lay a foundation for further studying the gene functions and screening the immunodiagnostic antigens.

Adult ; Diagnosis, Differential ; Dyspepsia ; blood ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Motilin ; blood

Objective To investigate the effect of pigment epithelium- derived factor (PEDF) on p38MAPK-CREB pathway and the expression of fibronectin (FN) in human mesangial cells (HMCs) cultured with high glucose. Methods HMCs were treated with different concentrations of glucose and the osmotic control respectively in the presence or absence of PEDF for 24 h:normal glucose (5.6 mmol/L),24.4 mmol/L mannitol,high glucose (30 mmol/L),high glucose+PEDF(30 mmol/L glucose with 10 nmol/L PEDF,40 nmol/L PEDF or 100 nmol/L PEDF).After samples were collected,the expression of phospho-p38MAPK (p-p38) and p-CREB was assessed by Western blotting,while FN mRNA and protein expression was assessed with RT-PCR and ELISA respectively. Results In contrast to normal glucose and mannitol treatments,the expression of p-p38MAPK,p-CREB and FN increased significantly in high glucose group (all P＜ 0.01).However,PEDF abolished the up-regulation of p-p38MAPK,p-CREB and FN induced by high glucose (all P＜0.05). Conclusion PEDF may inhibit fibrosis through P38MAPK-CREB pathway in diabetic nephropathy.

Objective To retrospectively investigate the endoscopic characteristics and pathological features of colorectal polyps in over-aged patients (≥80 years). Methods The 1617 colonoscopies performed between January 2006 and December 2010 were enrolled in our retrospective analysis. The detection rate, size, location, form and pathological feature of polyps in 150 over-aged patients were investigated, and they were compared with those in 832 young patients (＜60 years) and 635 old patients (60-79 years) with colorectal polyps. Results The detection rate of polyps was 62.0% in over-aged group, and was significantly higher than in young group and old group (30.2% and 48.7%, respectively, χ2=56.58 and 8.64,both P＜0.001). The malignant transformation rate of ascending colon polyp was 5.4% in over-aged group, and was significantly higher than in young group (1.2%, χ2=4.90, P＜0.05), but there was no significant difference between over-aged group and old group (3.9%, χ2=0.36, P＞0.05). There were no statistical differences in canceration rate, polyp diameter, morphology and adenomatous polyp rate among the three groups. The malignant transformation rate was significantly increased in adenomatous polyps with diameter over 2 cm, but didn't reach statistical significance. Conclusions The polyp detection rate is noticeably higher in the over-aged than in the old and the middle-aged, and the malignant transformation probability is also increased. The colonoscopy indications in the high risk age groups should appropriately be broadened, they should receive regular intervals follow-up, and undergo polypectomy in time if necessary.

Objective To discuss the relationship between serum 25-hydroxy vitamin D[25(OH)D] level and the severity of diabetic retinopathy in type 2 diabetes mellitus.Methods 1474 cases of type 2 diabetic patients were classified into three groups according to their severity of diabetic retinopathy, the non-diabetic retinopathy group(NDR group, n =640), the background diabetic retinopathy group(BDR group, n =494) and the proliferative diabetic retinopathy group(PDR group, n=340).640 healthy people with normal level of glycosylated hemoglobin(HbA1c) were selected as control group(NDM group, n=640).Disease duration, body mass index(BMI), systolic pressure, low density lipoprotein-cholesterol(LDL-C), HbA1c and 25(OH)D were detected and compared among groups.Results Disease duration and HbA1c were increasing significantly, and the levels were NDM groupNDR group>BDR group>PDR group(all P<0.05).The level of 25(OH)D was negatively related to disease duration and HbA1c (P<0.001), and it was negatively associated to the severity of diabetic retinopathy(r=-0.855,P<0.001).Conclusion The decrease of 25(OH)D level is closely associated with the severity of diabetic retinopathy in patients with type 2 diabetes mellitus, and it might be an important predictor for the severity of diabetic retinopathy.

OBJECTIVE:To study the effects of podophyllotoxin derivative QW-83 on human cervical cancer HeLa cell apopto-sis and its mechanism. METHODS:After treated with 0(negative control),0.01,0.1,1 and 10 μmol/L QW-83 and positive drug etoposide(VP-16)for 48 h,proliferation inhibition rate and IC50 of HeLa cell were determined by MTT assay. The morphological changes of HeLa cell were observed by Hochest 33342 staining after treated with QW-83 [0(negative control),2.5,5,10μmol/L] for 48 h;flow cytometry was used to detect apoptosis rate;semi quantitative RT-PCR was adopted to detect the expression of apop-tosis related gene P53,Bax,Casepase-3,Casepase-8,Casepase-9 and Bcl-2 mRNA. RESULTS:Compared with negative control, 1,10 μmol/L VP-16 and QW-83 had obvious proliferation inhibition effect on HeLa cells (P<0.05 or P<0.01),and IC50 were (5.11±0.43)μmol/L and(4.96±0.54)μmol/L. Hochest 33342 staining results showed QW-83 could obviously induce cells apopto-sis and nuclear pyknosis. Flow cytometry showed QW-83 could increase apoptosis rate in concentration-dependent manner,being 16.89%-62.56%. RT-PCR showed mRNA expression of P53,Bax,Caspase-3,Casepase-8 and Casepase-9,Bcl-2/Bax increased, while mRNA expression of Bcl-2 decreased after treated with QW-83(P<0.05). CONCLUSIONS:Podophyllotoxin derivative QW-83 can induce HeLa cell apoptosis,and its mechanism may be associated with regulate mRNA expression of apoptosis related gene.