Objective:To construct an adenovirus-mediated anti-sense RNA targeting K-ras exon 1 of SW1990 cell line and observe its effect on ceil proliferation and apoptosis after transferred into SW1990 cell line.Methods:K-ras exon 1 cDNA was cloned into shuttle vector pShuttle-CMV and the resultant plasmid was confirmed by enzyme digestion and PCR.Clones with inverted insertion were selected and co-transferred into E.coli BJ5183 with an adenoviral backbone plasmid pAdEasy-1 to produce recombinant plasmid by homologous recombination.Recombinants were then selected and transfected into 293 cell line to produce recombinant adenovirus.Recombinant adenovirus production was confirmed by PCR analysis and was amplified and purified;the virus titer was determined.Ad-LacZ was used to infect SW1990 cells and the infection efficiency was observed by X-gal staining.SW1990 cells was infected with the recombinant adenovirus and their proliferation and apoptosis were determined by MTT and annexin V/PI FCM assay.Results:A 282 bp target gene fragment was acquired by PCR;the titer of recombinant adenovirus was 7.6?10~8 pfu/ml before purification by CsCl_2 gradient centrifugation and 5.0?10~(10)pfu/ml after CsCl_2 gradient centrifugation.When the recombinant adenovirus was at 100 MOI,the infection efficiency of SW1990 cells nearly reached 100%.The transfection of recombinant adenovirus significantly inhibited SW1990 cell proliferation(P

OBJECTIVETo explore the relationship between the genesis of Pi-Wei damp-heat syndrome (PDS) and the Helicobacter pylori (HP) infection in patients with chronic gastritis (CG) by observing the levels of nuclear factor kappaB (NF-kappaB) mRNA and heat shock protein 70 (HSP70) expressions.

METHODSGastric mucous membrane tissues collected through gastroscopy from gastric antrum of 51 CG patients, 36 of PDS type (CG-PDS) and 15 of Pi-qi deficiency syndrome (CG-non-PDS) type, and 8 healthy persons (as control) were examined to diagnose the inflammation level with HE stain and to detect the existence of HP infection by rapid urease test and methylthioninium chloride stain. The mRNA expressions of NF-kappaB and HSP70 in the tissues were determined quantitatively with FQ-PCR as well.

RESULTSPatients of CG-PDS showed higher level of HSP70 mRNA expression than in the control; and level of NF-kappaB mRNA expression was higher than that in the control and CG-non-PDS (all P<0.05); but both expressions were insignificantly different in CG-PDS patients with positive or negative HP infection (P>0.05). Positive correlation of the two expressions was shown in CG-PDS with negative HP infection.

CONCLUSIONNF-kappaB and HSP70 may partially embody, in some extent, the power of vital qi and evil qi in the organism; the over-expressed NF-kappaB and HSP70 may indicate the severe fighting between evil qi and vital qi, and both would be influenced to a certain degree in the fighting process.

Adult ; Case-Control Studies ; Female ; Gastric Mucosa ; metabolism ; Gastritis ; diagnosis ; metabolism ; microbiology ; HSP70 Heat-Shock Proteins ; genetics ; metabolism ; Helicobacter Infections ; metabolism ; Helicobacter pylori ; Humans ; Male ; Medicine, Chinese Traditional ; NF-kappa B ; genetics ; metabolism ; RNA, Messenger ; genetics

We constructed plasmid pMTac to overexpress 3-ketosteroid-Δ1-dehydrogenase (KSDD) in Mycobacterium neoaurum JC-12 for improving androst-1,4-diene-3,17-dione (ADD) production. To construct pMTac, pACE promoter on pMF41 was replaced by tac promoter, and then four recombinants were constructed, which were M. neoaurum JC-12/pMF41-gfp, M. neoaurum JC-12/pMTac-gfp, M. neoaurum JC-12/pMF41-ksdd and M. neoaurum JC-12/pMTac-ksdd. Fluorescence detection results show that much more green fluorescent protein (GFP) was expressed in M. neoaurum JC-12/pMTac-ksdd than M. neoaurum JC-12/pMF41-ksdd. The activity of KSDD was 2.41 U/mg in M. neoaurum JC-12/pMTac-ksdd, 6.53-fold as that of M. neoaurum JC-12 and 4.36-fold as that of M. neoaurum JC-12/pMF41-ksdd. In shake flask fermentation, ADD production of M. neoaurum JC-12/pMTac-ksdd was 5.94 g/L, increased about 22.2% compared to the original strain M. neoaurum JC-12 and 12.7% to M. neoaurum JC-12/pMF41-ksdd. AD (4-androstene-3,17-dione) production of JC-12/pMTac-ksdd was 0.17 g/L, decreased 81.5% compared to M. neoaurum JC-12 and 71.2% to M neoaurum JC-12/pMF41-ksdd. In the 5 L fermenter, 20 g/L phytosterols was used as substrate, ADD production of M. neoaurum JC-12/pMTac-ksdd was improved to 10.28 g/L. pMTac is favorable for expressing KSDD in M. neoaurum JC-12, and overexpression of KSDD has beneficial effect on ADD producing, and it is the highest level ever reported using fermentation method in M. neoaurum.
Androstadienes ; metabolism ; Fermentation ; Industrial Microbiology ; Mycobacterium ; Oxidoreductases ; genetics ; metabolism ; Phytosterols ; metabolism ; Plasmids

Objective To study the clinical significance of the serum angiopoietin-2(Ang-2) in the diagnosis,recurrence,invasion and metastasis of gastric cancer.Methods The serum Ang-2 and carcino-embryonic antigen (CEA) levels in 158 patients with gastric cancer (gastric cancer group) and 30 normal controls(control group) were measured by enzyme linked immunosorbent assay(ELISA) technique respectively.The serum Ang-2 and CEA levels were also measured 2 weeks after operation in gastric cancer group and reexamined in the recurred gastric cancer patients in 2 years after operation (recurred and metastasis group).The correlation between the serum Ang-2 level and pathologic c haracterization of gastric cancer was evaluated.Results The serum Ang-2 and CEA levels in gastric cancer group [ (331.8 ±64.3),(42.6 ±37.3)μg/L] and recurred and metastasis group [(318.7 ±72.9),(40.5 ±36.7)μg/L] were significantly higher than those in control group [ (187.4 ± 32.7),(4.2 ± 3.1 )μ g/L] (P ＜ 0.01 ),and the serum Ang-2 level 2 weeks after operation [ (211.6 ± 75.1 ) μ g/L ] was significantly decreased to the control group (P ＞ 0.05 ),while the serum CEA level [ (33.4 ± 30.6) μ g/L ] was still significantly higher than the control group (P ＜ 0.01 ).The sensitivity of the serum Ang-2 for diagnosis of gastric cancer was markedly higher than that of the serum CEA (P ＜ 0.01 ).There was correlation between serum Ang-2 and degree of tumor differentiation,TNM pathological staging,lymphatic metastasis,invasion depth and tumor size (p ＜0.01 ),but there was no correlation between serum Ang-2 and tissue classification and location of gastric cancer (P＞ 0.05).Conclusion The serum Ang-2 level is suggested to be a valuable gastric cancer marker and conduce to the diagnosis of gastric cancer,the monitoring of recurrence after operation and evaluation of prognosis.

Objective To explore the quality of life (QOL) and its influential factors among patients with 3 major rheumatic diseases. Methods A total of 216 patients with rheumatic diseases (84 patients with systemic lu- pus erythematosus, SLE, 83 with rheumatoid arthritis, RA, and 49 with ankylosing spondylitis, AS) were recruited. The information with regard to their quality of life, sociopsychological factors and the evaluation of disease activity were obtained by using the medical outcomes study short form-36 (SF-36) and clinic documents. Results Patients with rheumatic diseases scored significantly lower with each subscale of SF-36 as compared to those of a healthy popu- lation in China (P

Objective:To search for a method for radical resection of pancreatic and duodenal malignancy involving the mesentery root and for the long post-operation survival of patients.Methods:From Jan.2004 to Aug.2006,a total of 26(16 male and 10 female. aged 27-70)patients with pancreatic and duodenal malignancy involving the mesentery root were treated in our department.The patients included 3 with duodenal malignancy and 23 with pancreatic malignancy.Curative resection was performed by the extended pancreaticoduodenetomy(Whipple procedure)combined with mesentery root resection(MRR)for all patients.The outcomes,safety and the post-operation survival rate were analyzed retrospectively.Results:Thirteen patients were treated with Whipple procedures combined with MRR,9 were treated with partial portal vein/superior mesenteric vein(PV/SMV)and reconstruction of the vessel,and 4 patients received pre-shunt between PV and SMV with artificial vessel graft before the extended Whipple and MRR procedures.The operation time was 2.5 to 7(4.4?1.1)hour,and blood loss was 300 to 5 000(1892?1414)ml with the blood transfusion of 0 to 5 600(2 100?1 586)ml.There was no death in our group and 7(27%)had post-operation complication.The post-operation hospital stay was 10 to 30 days.The pathologic examination showed negative surgical margins for all specimens.The tumor size was 4 to 10 (6.17?2.03)cm.After a follow-up of 9 to 38 months,the pain was relieved in all patients.One of the 3 patients with duodenal adenocarcinoma had liver metastasis at 10 months after operation,and the other 2 survived 10 months and 27 months without evidence of tumor reccurence.The patient with pancreatic micro-adenocarcinoma died of local reccurence 9 months after operation.The patient with neuroendocrine carcinoma died of organ failure 24 months after operation.The patient with lymphoma have survived for 24 months after operation.The 1-year and 2-year accumulated survival rates in the 20 cases with pancreatic ductal cancer were 86.6% and 45.6%. respectively.Conclusion:The extended Whipple procedure with MRR is safe and effective.It can obtain R0 resection in patients with malignant tumors(over 5 cm in diameter)in the head,neck and uncinate process of the pancreas and duodenal.

It has been proved in recent studies that the chronic gastric disease (CGD) of Pi-Wei damp-heat syndrome type (CGD-PWDH) is closely related with heat shock protein 70 (HSP70) and nuclear factor-kappa B (NF-kappaB). HSP70 can protect the auto-stability of cells and elevate the immune function in organism against tumor or multiple exogenous pathogens. Increasing of NF-kappaB expression presents in case of Helicobacter pylori (Hp) stimulation, it could induce inflammatory reaction, while inflammation factors could act inversely to enhance the expression of NF-kappaB, thus to cause and expand the damage of gastric mucosa. In addition, HSP shows blocking effect on the activation and expression of NF-kappaB. So, the author considered that in patients of Hp associated CGD-PEDH, HSP 70 exhibits the effect as that of "vital energy" and NF-kappaB play a role as the "evil qi" in Chinese medicine, the expressions of the two may embody the vital-evil combating manner of Pi-Wei damp-heat syndrome.
Gastrointestinal Diseases ; diagnosis ; metabolism ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Medicine, Chinese Traditional ; methods ; NF-kappa B ; metabolism

Pi-Wei theory is an important component of the basic theory of Chinese medicine. The pathogenesis of Pi-Wei damp-heat syndrome is an important content of Pi-Wei theory. The tongue coating is one of the most important signs reflecting Pi-Wei damp-heat syndrome. From the perspective of microecology and pathogenesis, the microbial disequilibrium caused by quantity changes of Helicobacter pylori (Hp) and Lactobacillus acidophilus (LA) and their interaction in the gastric mucosa and the tongue coating might have certain correlation with "mutual struggle between the evil and the vital qi, the disequilibrium between yin and yang". The pathogenesis features of chronic gastritis patients of Pi-Wei damp-heat syndrome was initially proposed in this article.
Diagnosis, Differential ; Gastric Mucosa ; microbiology ; Helicobacter Infections ; diagnosis ; Helicobacter pylori ; isolation & purification ; Humans ; Lactobacillus acidophilus ; isolation & purification ; Medicine, Chinese Traditional ; methods ; Stomach Diseases ; diagnosis ; microbiology

Adult ; Brain Neoplasms ; diagnosis ; secondary ; Female ; Humans ; Lymphomatoid Granulomatosis ; complications ; diagnosis ; Male ; Young Adult

The purpose of the present study was to investigate the effect of 17beta-estradiol (17beta-E(2)) on the structure and relaxation and contraction activity of thoracic aortas in ovariectomized rats with insulin resistance induced by fructose. Ovariectomized mature female Sprague-Dawley rats were fed with high fructose diet for 8 weeks to induce insulin resistance. Physiological dose of 17beta-E(2) (30 mug/kg) was injected subcutaneously every day for 8 weeks. Systolic blood pressure (SBP) was measured by use of tail-cuff. Serum nitric oxide (NO), estradiol (E(2)), fasting blood sugar (FBS) and fasting serum insulin (FSI) were measured respectively in each group. The insulin sensitive index (ISI) was calculated. The thoracic aortas were fixed in formalin, sliced and HE dyed. The structure of thoracic aortas, lumen breadth, media thickness, media thickness/lumen breadth ratio and media cross-section area were measured. The contraction response of thoracic aorta rings induced by L-phenylephrine (PE) and the relaxation response of thoracic aorta rings induced by ACh and sodium nitroprusside (SNP) were measured. To explore the mechanism, nitric oxide synthase (NOS) inhibitor N-nitro-L-arginine methyl ester (L-NAME) was used. The results obtained are as follows: (1) 17beta-E(2) protected against the effect of high fructose diet, which caused an increase in SBP, hyperinsulinemia and a decrease in ISI in ovariectomized rats. (2) The structure of thoracic aortas had no significant difference among the groups. (3) Compared with the ovariectomized group (OVX) or fructose fed group (F), serum nitric oxide was significantly reduced, the contraction response of thoracic aorta rings to PE was enhanced and the relaxation response to ACh was depressed significantly in ovariectomized+fructose fed group (OVX+F). The effect of high fructose was reversed by 17beta-E(2). After pretreatment with L-NAME, the effect of 17beta-E(2), which enhanced the relaxation response of thoracic aorta rings to ACh in ovariectomized+fructose+17beta-E(2) group (OVX+F+E(2)), was partly blocked. (4) The relaxation response of thoracic aorta rings to SNP had no significant difference among the groups. (5) The contraction response of thoracic aorta rings without endothelium to PE had no significant difference among the groups. These findings suggest that 17beta-E(2) may provide protection against the effect of high fructose diet, which causes hypertension, dysfunction of endothelial cells and insulin resistance. The mechanism of this effect of 17beta-E(2) could be partly associated with the increase of NO by NOS pathway, or associated with the decrease in the level of systolic blood pressure and serum insulin, and the improvement of insulin resistance.
Animals ; Aorta ; physiology ; Estradiol ; pharmacology ; Female ; Fructose ; Insulin Resistance ; physiology ; Ovariectomy ; Rats ; Rats, Sprague-Dawley ; Vasoconstriction ; drug effects ; Vasodilation ; drug effects ; Vasomotor System ; drug effects